MH_dev_42

Molecular weight and biochemical profile of a chemically modified heparin derivative , Suleparoide . Recently , a new chemically modified derivative of heparin ( Suleparoide , Syntex Laboratories Buenos Aires , Argentina ) was introduced for the prophylaxis of thrombosis and treatment of vascular disorders . This agent is claimed to contain a depolymerized , chemically modified , heparin derivative with similar biologic actions as heparan sulfate . To study the pharmacologic profile of this agent , we have defined its molecular weight distribution profile , utilizing a computerized gel permeation chromatographic system equipped with ultraviolet and refractive index detectors . Suleparoide exhibited a normal molecular distribution profile with no contaminants . It exhibited a weight average of 9.3 K DA and an apparent peak MW of 8.0 K DA . Approximately 50 % of the molecular components were < 5.0 K DA and 40 % > 5.0 K DA . The results from these studies on the mechanisms show that Suleparoide has anticoagulant activity primarily mediated through DB01109 MEN Cofactor-II ( P05546 REA ) and because of its novel mechanism of action , further investigations on the biochemical profile of Suleparoide are carried out . Global clotting tests such as Activated Partial P13726 REA Time ( APTT ) , Heptest and Thrombin Time ( TT ) revealed a concentration dependent effect in all assays . Plasma samples supplemented with Suleparoide exhibited no significant anti-Xa and anti-IIa activities . However , in the P05546 REA mediated inhibitory assay for IIa , Suleparoide exhibited significant activity . In contrast , the P01008 REA ( DB11598 ) mediated inhibition of IIa was much weaker .

Copy number analysis of 24 oncogenes : O15151 REA identified as a putative marker for low recurrence risk in non muscle invasive bladder cancer . Patients with non-muscle invasive bladder cancer ( NMIBC ) generally have a high risk of relapsing locally after primary tumor resection . The search for new predictive markers of local recurrence thus represents an important goal for the management of this disease . We studied the copy number variations ( CNVs ) of 24 oncogenes ( O15151 REA , P04198 , Q9UM73 , P16234 REA , P10721 REA , P35968 REA , P00374 REA , P00533 REA , MET , SMO , P11362 REA , MYC , P00519 REA , P07949 REA , P24385 REA , P30279 REA , P11802 REA , Q00987 REA , Q96GD4 , P04626 REA , P11388 REA , O14965 REA , AR and P15056 REA ) using multiplex ligation probe amplification technique to verify their role as predictive markers of recurrence . DB03843 - fixed paraffin-embedded tissue samples from 43 patients who underwent transurethral resection of the bladder ( TURB ) were used ; 23 patients had relapsed and 20 were disease-free after 5 years . Amplification frequencies were analyzed for all genes and O15151 REA was the only gene that showed significantly higher amplification in non recurrent patients than in recurrent ones ( 0.65 vs . 0.3 ; Fisher ' s test p= 0.023 ) . Recurrence-free survival analysis confirmed the predictive role of O15151 REA ( log-rank test p= 0.041 ) . Our preliminary results indicate a putative role for the O15151 REA gene in predicting local recurrence of bladder cancer . Confirmation of this hypothesis is needed in a larger cohort of NMIBC patients .

[ Bone metastasis in prostate cancer ] . Bone metastasis and skeletal complications have a devastating impact on the quality of life and are a major cause of morbidity in prostate cancer patients . In addition to established bone-targeted therapies , new drugs such as endothelin A receptor antagonists , MET and P35968 REA antagonists or radiopharmaceuticals are in the focus of development . The standard care in prostate cancer patients with bone metastases to prevent skeletal-related events ( SRE ) are bisphosphonates . DB06643 MEN , a human monoclonal antibody against O14788 REA , appeared to be superior to zoledronic acid for prevention of SRE and has been shown to prolong bone metastases-free survival . In contrast to zoledronic acid , denosumab clearance is not dependent on kidney function and can be administered subcutaneously . Similar rates of toxicity were observed for both substances ; however , long-term data for denosumab are limited .

A valuable adjunct to FNA diagnosis of papillary thyroid carcinoma : in-house PCR assay for P15056 REA T1799A ( V600E ) . The diagnostic approach to thyroid nodules generally starts with FNA cytology . However , approximately one-fifth of cytologic evaluations yield indeterminate cytological findings but only 20 % of cases with indeterminate thyroid nodule cytology have a cancer diagnosis , emphasizing the need for an effective ancillary test based on FNA material to help prevent unnecessary surgery . Detection of BRAFV 600E mutation , the genetic signature of papillary thyroid carcinoma ( PTC ) in FNA material provides an invaluable diagnostic adjunct to overcome the limitations of FNA cytology . There are many ways to detect V600E , such as direct DNA sequencing , allele-specific PCR and hybridization-based colorimetric methods . In this study , a newer simple PCR method is presented that removes requirements for sequencing special equipment and commercial kits . Two forward primers including the mutant sequence specific ( F2 ) , and one common reverse ( R ) primer were optimized to generate a 241 bp fragment ( F1R ) , an internal PCR control , and a 141 bp fragment ( P25116 REA ) denoting the presence of V600E . Sensitivity studies revealed that the assay is capable of detecting V600E even in 1 ng of DNA . Direct sequencing data of 241 bp F1R fragment proved the specificity of the assay . For validation studies of the sequence specific multiplex PCR assay , archival FNA slides were used in a group of thyroid lesions including PTC , follicular carcinoma , follicular adenoma , Hashimoto thyroiditis , and benign thyroid nodules . The newer PCR-based method presented in this study is a practical , inexpensive one-step assay to detect the P15056 REA T1796A mutation on FNA samples .

DB00741 MEN is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 MEN ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 REA ) and caspase 3 ( P42574 REA ) and reduced the enzymatic activity of P42574 REA and cell death induced by tumor necrosis factor ( P01375 REA ) and interferon gamma ( P01579 REA ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 REA ) , 11beta - hydroxysteroid dehydrogenase type 1 ( P28845 REA ) , and P8 0365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 REA were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 REA - P01579 REA - induced apoptosis in vitro by reducing apoptosis signals via Q14790 REA and P42574 REA in bovine CL and that the local increase in cortisol production resulting from increased P28845 REA at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells .

P10275 REA promotes esophageal cancer cell migration and proliferation via matrix metalloproteinase 2 . Esophageal squamous cell carcinoma ( ESCC ) is one of the most common malignancies worldwide . P10275 REA ( AR ) plays an important role in many kinds of cancers . However , the molecular mechanisms of AR in ESCC are poorly characterized . In the present study , Western blot analysis and real-time quantitative PCR were performed to identify differentially expressed AR in 40 ESCC tissue samples , which revealed that the messenger RNA ( mRNA ) and protein expression of AR is upregulated in the ESCC tissue samples . AR overexpression induced increases in ESCC cell invasion and proliferation in vitro . Silencing of AR inhibited the proliferation of KYSE 450 cells which have a relatively high level of AR , and the invasion of KYSE 450 cells was distinctly suppressed . Furthermore , AR knockdown led to substantial reductions in matrix metalloproteinase 2 ( P08253 REA ) and p-AKT levels in ESCC cell lines , but no significant change in AKT and P14780 REA expression . These results suggest that AR is involved in tumor progression , and thus , AR could represent selective targets for the molecularly targeted treatments of ESCC .

Molecular mechanisms and modulation of key pathways underlying the synergistic interaction of sorafenib with erlotinib in non-small-cell-lung cancer ( NSCLC ) cells . Combination of drugs with different targets is a logical approach to overcome multilevel cross-stimulation among key pathways in NSCLC progression such as P00533 REA , K-Ras and VEGFR . The sorafenib-erlotinib combination showed clinical activity and acceptable safety . Therefore , we evaluated mechanisms underlying sorafenib-erlotinib interaction in seven NSCLC cell lines selected for their heterogeneous pattern of P00533 REA and Raf-kinase-inhibitor protein ( P30086 REA ) expression , and P00533 REA / K-Ras mutations . Pharmacologic interaction was studied using MTT / P50991 REA assays and the combination index ( CI ) method , while effects on P00533 REA , Erk 1/2 and Akt phosphorylation , cell cycle and apoptosis were studied with western-blot , ELISA , and flow cytometry . Intracellular drug concentrations were measured with LC-MS / MS , whereas kinase activity profiles were generated on tyrosine kinase peptide substrate arrays . Synergism was detected in all cell lines , with CIs < 0.6 in K-Ras mutated A549 , SW1573 and H460 , as well as in H1975 ( P00533 REA - T790M ) cells . DB00398 SUB slowed cell cycle progression and induced apoptosis , which was significantly increased in the combination . Moreover , sorafenib reduced Akt / P29323 REA phosphorylation in erlotinib-resistant cells , associated with significant P30086 REA up-regulation . No direct drug interaction was detected by LC-MS / MS measurement , while lysates from A549 and H1975 cells exposed to erlotinib + sorafenib showed a significant inhibition in the phosphorylation of 16 overlapping peptides , including sites from RAF , P35968 REA , P09619 REA , P24941 REA and P12931 REA , suggesting new markers to identify NSCLC patients who are likely to respond to this treatment . In conclusion , several mechanisms , including apoptosis-induction , modulation of expression / phosphorylation of P30086 REA and crucial kinases contribute to erlotinib-sorafenib synergistic interaction and should be evaluated in future trials for the rational development of this combination in NSCLC .

Temporal and pharmacological characterization of angiostatin release and generation by human platelets : implications for endothelial cell migration . Platelets play an important role in thrombosis and in neo-vascularisation as they release and produce factors that both promote and suppress angiogenesis . Amongst these factors is the angiogenesis inhibitor angiostatin , which is released during thrombus formation . The impact of anti-thrombotic agents and the kinetics of platelet angiostatin release are unknown . Hence , our objectives were to characterize platelet angiostatin release temporally and pharmacologically and to determine how angiostatin release influences endothelial cell migration , an early stage of angiogenesis . We hypothesized anti-platelet agents would suppress angiostatin release but not generation by platelets . Human platelets were aggregated and temporal angiostatin release was compared to vascular endothelial growth factor ( P15692 REA ) . Immuno-gold electron microscopy and immunofluorescence microscopy identified α-granules as storage organelles of platelet angiostatin . Acetylsalicylic acid , MRS 2395 , P08514 REA / IIIa blocking peptide , and aprotinin were used to characterize platelet angiostatin release and generation . An endothelial cell migration assay was performed under hypoxic conditions to determine the effects of pharmacological platelet and angiostatin inhibition . Compared to P15692 REA , angiostatin generation and release from α-granules occurred later temporally during platelet aggregation . Consequently , collagen-activated platelet releasates stimulated endothelial cell migration more potently than maximally-aggregated platelets . Platelet inhibitors prostacyclin , S-nitroso-glutathione , acetylsalicylic acid , and P08514 REA / IIIa blocking peptide , but not a Q9H244 REA inhibitor , suppressed angiostatin release but not generation . Suppression of angiostatin generation in the presence of acetylsalicylic acid enhanced platelet-stimulated endothelial migration . Hence , the temporal and pharmacological modulation of platelet angiostatin release may have significant consequences for neo-vascularization following thrombus formation .

Clinical outcome , role of P15056 REA ( V600E ) , and molecular pathways in papillary thyroid microcarcinoma : is it an indolent cancer or an early stage of papillary thyroid cancer ? Most human thyroid cancers are differentiated papillary carcinomas ( PTC ) . Papillary thyroid microcarcinomas ( PTMC ) are tumors that measure 1 cm or less . This class of small tumors has proven to be a very common clinical entity in endocrine diseases . PTMC may be present in 30-40 % of human autopsies and is often identified incidentally in a thyroid removed for benign clinical nodules . Although PTMC usually has an excellent long-term prognosis , it can metastasize to neck lymph nodes ; however deaths related to this type of thyroid tumor are very rare . Few data exist on molecular pathways that play a role in PTMC development ; however , two molecules have been shown to be associated with aggressive PTMC . P26447 ( calcium-binding protein ) , which plays a role in angiogenesis , extracellular matrix remodeling , and tumor microenvironment , is over-expressed in metastatic PTMC . In addition , the P15056 REA ( V600E ) mutation , the most common genetic alteration in PTC , is present in many PTMC with extra thyroidal extension and lymph node metastasis . Importantly , recently developed selective [ e . g . , PLX 4720 , PLX 4032 ( DB08881 , also called RG7204 ) ] or non-selective ( e . g . , DB00398 SUB ) inhibitors of P15056 REA ( V600E ) may be an effective treatment for patients with P15056 REA ( V600E ) - expressing PTMCs with aggressive clinical-pathologic features . Here , we summarize the clinical outcome , cancer genetics , and molecular mechanisms of PTMC .

DB00398 SUB inhibits imatinib-resistant P10721 REA and platelet-derived growth factor receptor beta gatekeeper mutants . PURPOSE : Targeting of P10721 REA and platelet-derived growth factor receptor ( P09619 REA ) tyrosine kinases by imatinib is an effective anticancer strategy . However , mutations of the gatekeeper residue ( T670 in P10721 REA and T681 in PDGFRbeta ) render the two kinases resistant to imatinib . The aim of this study was to evaluate whether sorafenib ( BAY 43-9006 ) , a multitargeted DB00171 - competitive inhibitor of P10721 REA and P09619 REA , was active against imatinib-resistant P10721 REA and PDGFRbeta kinases . EXPERIMENTAL DESIGN : We used in vitro kinase assays and immunoblot with phosphospecific antibodies to determine the activity of sorafenib on P10721 REA and PDGFRbeta kinases . We also exploited reporter luciferase assays to measure the effects of sorafenib on P10721 REA and PDGFRbeta downstream signaling events . The activity of sorafenib on interleukin - 3 - independent proliferation of Ba / P13726 REA cells expressing oncogenic P10721 REA or its imatinib-resistant T670I mutant was also tested . RESULTS : DB00398 SUB efficiently inhibited gatekeeper mutants of P10721 REA and PDGFRbeta ( IC ( 50 ) for P10721 REA T670I , 60 nmol / L ; IC ( 50 ) for PDGFRbeta T681I , 110 nmol / L ) . Instead , it was less active against activation loop mutants of the two receptors ( IC ( 50 ) for P10721 REA D8 16V , 3.8 micromol / L ; IC ( 50 ) for PDGFRbeta D8 50V , 1.17 micromol / L ) that are also imatinib-resistant . DB00398 SUB blocked receptor autophosphorylation and signaling of P10721 REA and PDGFRbeta gatekeeper mutants in intact cells as well as activation of P05412 REA - responsive and cyclin D1 gene promoters , respectively . Finally , the compound inhibited P10721 REA - dependent proliferation of Ba / P13726 REA cells expressing the oncogenic P10721 REA mutant carrying the T670I mutation . CONCLUSIONS : DB00398 SUB might be a promising anticancer agent for patients carrying P10721 REA and PDGFRbeta gatekeeper mutations .

Potential role of activated platelets in homing of human endothelial progenitor cells to subendothelial matrix . Endothelial progenitor cells ( EPCs ) mobilize from the bone marrow in response to tissue injury and participate in vascular repair . However , there is limited data about the homing mechanisms of EPCs to vascular injury sites . Recently animal experiments indicated that platelets play a role in recruitment of EPCs to injury sites . However , data on the possible interaction between platelets and EPCs within the human system are limited . We , therefore , examined in-vitro human platelet-EPC interaction under static and flow conditions . Human EPCs were isolated from donated buffy coats by magnetic microbeads and flow cytometry cell sorting using CD133 and P35968 REA , respectively , as markers . Platelets were tested in the form of washed platelets , platelet rich plasma or whole blood . EPCs formed heterotypic aggregates with resting platelets under static conditions , an interaction that was greatly enhanced when platelets were activated by collagen , ADP or thrombin-activation peptide . The platelet-EPC interaction was inhibited by antibodies to P16109 REA or P16109 REA glycoprotein ligand - 1 ( Q14242 REA ) , but not by antibodies to glycoproteins Ib-IX-V or IIb / IIIa . When perfused over activated platelets under shear stress of 2.5 dyn / cm ( 2 ) , EPCs tethered to platelayers and either adhered immediately or rolled a short distance before adhering . In addition , platelets promoted the colonization of adherent EPCs in culture conditions . Consistent with recent animal studies , these findings demonstrate that human EPCs interact in vitro with activated platelets under static and flow conditions , mediated through P16109 REA - Q14242 REA interaction . This interaction may be a central mechanism for homing of EPCs to vascular injury sites .

Order of genes on human chromosome 5q with respect to 5q interstitial deletions . Using ( a ) somatic cell hybrids retaining partial chromosome 5 and ( b ) clinical samples from patients with acquired deletions of the long arm of chromosome 5 , combined with chromosome 5 - linked DNA probes , some of which exhibited RFLPs , we have determined the order of a series of genes on chromosome 5 . The order established is 5pter - - - MLVI - 2 - - - cen - - - P07686 REA - - - P00374 REA - - - Pi227 - - - - cp12 . 6 - - - ( P05113 REA , P05112 REA ) - - - P08700 REA - - - P04141 REA - - - P05230 REA - - - ( P07333 REA , P09619 REA ) - - - ( treC , ADRBR ) - - - ( Q5SW96 - Q13585 , P09603 REA ) - - - qter . The suggested order and orientation for the closely linked P08700 REA / P04141 REA gene pair is cen - - - 5 ' P08700 REA 3 ' - - - 5 ' P04141 REA 3 ' - - - qter , on the basis of analysis of the P04141 REA rearrangement in HL60 DNA . The map position of the GRL locus , which was consistent with both somatic cell hybrid and 5q - analyses , was telomeric to P04141 REA and centromeric to P07333 REA / P09619 REA , near P05230 REA . Long-range restriction-enzyme analysis of 5q - DNAs did not detect rearrangements of 5q - linked probes except in HL60 DNA , but it did reveal putative long-range RFLPs of several loci . RFLPs for GRL , Pi227 , cp12 . 6 , P08700 REA , and P07333 REA can detect deletions in bone marrow and in leukemia cells from patients with acquired 5q deletions .

Toxicity of sorafenib : clinical and molecular aspects . IMPORTANCE OF THE FIELD : DB00398 SUB is a novel oral bis-aryl urea compound originally developed as an inhibitor of RAF kinase for its anti-proliferative property . DB00398 SUB also inhibits receptor tyrosine kinases of multiple pro-angiogenic factors such as P17948 REA / 2/3 , Flt - 3 and P09619 REA . The combination of both its anti-proliferative and anti-angiogenic properties makes sorafenib an attractive agent in cancer treatment . DB00398 SUB has been approved for the treatment of metastatic renal cell carcinoma as well as hepatocellular cancer . Despite its inherent selectivity , sorafenib can cause unusual adverse events whose the management represents a challenge for oncologists . AREAS COVERED IN THIS REVIEW : Relevant literature was identified using a Pubmed search of articles published up to June 2009 . Search terms included ' sorafenib ' and ' toxicity ' . Original articles were reviewed and relevant citations from these articles were also considered . WHAT THE READER WILL GAIN : The clinical aspect of sorafenib-induced adverse events and the molecular basis behind this toxicity are discussed . Finally , recommendations for the management of these adverse events are proposed . TAKE HOME MESSAGE : Although not life-threatening , toxicity of sorafenib can severely impact the physical , psychological and social well-being of patients . The management of this unusual toxicity highlights the particular need of new pluridisciplinarities linking oncologist , cardiologist and dermatologist .

M4 and M5 acute myeloid leukaemias display a high sensitivity to DB00188 - mediated apoptosis . The present study explored the sensitivity of leukaemic blasts derived from 30 acute myeloid leukaemia ( AML ) patients to DB00188 . DB00188 induced apoptosis of primary AML blasts : 18/30 AMLs were clearly sensitive to the proapoptotic effects of DB00188 , while the remaining cases were moderately sensitive to this molecule . The addition of tumour necrosis factor-related-apoptosis-inducing ligand , when used alone , did not induce apoptosis of AML blasts and further potentiated the cytotoxic effects of DB00188 . The majority of AMLs sensitive to DB00188 showed immunophenotypic features of the M4 and M5 French-American-British classification subtypes and displayed myelomonocytic features . All AMLs with mutated P36888 REA were in the DB00188 - sensitive group . Biochemical studies showed that : ( i ) DB00188 activated caspase - 8 and caspase - 3 and decreased cellular Q14790 REA [ Fas-associated death domain ( Q13158 REA ) - like interleukin - 1beta - converting enzyme ] - inhibitory protein ( O15519 REA ) levels in AML blasts ; ( ii ) high O15519 REA levels in AML blasts were associated with low DB00188 sensitivity . Finally , analysis of the effects of DB00188 on leukaemic cells displaying high aldehyde dehydrogenase activity suggested that this drug induced in vitro killing of leukaemic stem cells . The findings of the present study , further support the development of DB00188 as an anti-leukaemic drug and provide simple tools to predict the sensitivity of AML cells to this drug .

DB00398 SUB treatment of P36888 REA - ITD ( + ) acute myeloid leukemia : favorable initial outcome and mechanisms of subsequent nonresponsiveness associated with the emergence of a D8 35 mutation . Internal tandem duplication ( ITD ) of the fms-related tyrosine kinase - 3 ( P36888 REA ) gene occurs in 30 % of acute myeloid leukemias ( AMLs ) and confers a poor prognosis . Thirteen relapsed or chemo-refractory P36888 REA - ITD ( + ) AML patients were treated with sorafenib ( 200-400 mg twice daily ) . Twelve patients showed clearance or near clearance of bone marrow myeloblasts after 27 ( range 21-84 ) days with evidence of differentiation of leukemia cells . The sorafenib response was lost in most patients after 72 ( range 54-287 ) days but the P36888 REA and downstream effectors remained suppressed . Gene expression profiling showed that leukemia cells that have become sorafenib resistant expressed several genes including P00352 REA , P52333 REA , and P51511 REA , whose functions were unknown in AML . Nonobese diabetic / severe combined immunodeficiency mice transplanted with leukemia cells from patients before and during sorafenib resistance recapitulated the clinical results . Both ITD and tyrosine kinase domain mutations at D8 35 were identified in leukemia initiating cells ( LICs ) from samples before sorafenib treatment . LICs bearing the D8 35 mutant have expanded during sorafenib treatment and dominated during the subsequent clinical resistance . These results suggest that sorafenib have selected more aggressive sorafenib-resistant subclones carrying both P36888 REA - ITD and D8 35 mutations , and might provide important leads to further improvement of treatment outcome with P36888 REA inhibitors .

Dopamine modulating drugs influence striatal ( + ) - [ 11C ] DTBZ binding in rats : Q05940 REA binding is sensitive to changes in vesicular dopamine concentration . Binding of ( + ) - [ 11C ] DTBZ ( dihydrotetrabenazine ) to the striatal vesicular monoamine transporter ( Q05940 REA ) is widely considered to be a stable marker of dopamine neurone integrity . However , we now find that specific binding of a tracer dose of ( + ) - [ 11C ] DTBZ is modestly increased in rat striatum following dopamine depletion with alpha-methyl-p-tyrosine ( alpha-MPT , + 14 % ) or DB01576 MEN ( d - P49418 REA , 20 mg / kg , + 12 % ) and decreased following dopamine elevation with gamma-hydroxybutyrate ( DB01440 , - 16 % ) or levodopa ( - 20 % ) . We suggest that in vivo ( + ) - [ 11C ] DTBZ binding in imaging studies is subject to competition by vesicular dopamine and , in this respect , is not a " stable " dopamine biomarker as is generally assumed .

[ Cytoprotective and antiangiogenic effects of the multikinase inhibitor sorafenib on human retinal pigmentepithelium ] . BACKGROUND : Cumulative light exposure is significantly associated with progression of age-related macular degeneration ( AMD ) . Inhibition of vascular endothelial growth factor A ( P15692 REA ) is the main target of current antiangiogenic treatment strategies for AMD . Previous reports indicated that sorafenib , an oral multikinase inhibitor , might have beneficial effects on exudative AMD . This study investigates the effects of sorafenib on light-induced overexpression of P15692 REA and its receptors P17948 REA and 2 in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : The effects of sorafenib on P17948 REA and 2 expression of primary human Q96AT9 cells was investigated by reverse transcription polymerase chain reaction ( RT-PCR ) , immunohistochemistry and western blotting . In addition , Q96AT9 cells were exposed to white light and incubated with sorafenib . Viability , expression of P15692 REA and its mRNA were determined by RT-PCR , immunohistochemistry , western blotting , and enzyme-linked immunosorbent assays . RESULTS : DB00398 SUB reduced P17948 REA and 2 expression of Q96AT9 cells . Light exposure decreased cell viability and increased expression and secretion of P15692 REA . These light-induced effects were significantly reduced when cells were treated with sorafenib at a dose of 1 µg / ml . CONCLUSION : The results show that sorafenib has promising properties as a potential antiangiogenic treatment for AMD .

Novel Q02750 REA mutation identified by mutational analysis of epidermal growth factor receptor signaling pathway genes in lung adenocarcinoma . Genetic lesions affecting a number of kinases and other elements within the epidermal growth factor receptor ( P00533 REA ) signaling pathway have been implicated in the pathogenesis of human non-small-cell lung cancer ( NSCLC ) . We performed mutational profiling of a large cohort of lung adenocarcinomas to uncover other potential somatic mutations in genes of this pathway that could contribute to lung tumorigenesis . We have identified in 2 of 207 primary lung tumors a somatic activating mutation in exon 2 of Q02750 REA ( i . e . , mitogen-activated protein kinase kinase 1 or Q02750 REA ) that substitutes asparagine for lysine at amino acid 57 ( K57N ) in the nonkinase portion of the kinase . Neither of these two tumors harbored known mutations in other genes encoding components of the P00533 REA signaling pathway ( i . e . , P00533 REA , P04626 REA , P01116 REA , P42336 REA , and P15056 REA ) . Expression of mutant , but not wild-type , Q02750 REA leads to constitutive activity of extracellular signal-regulated kinase ( P29323 REA ) -1/2 in human 293T cells and to growth factor-independent proliferation of murine Ba / P13726 REA cells . A selective MEK inhibitor , AZD 6244 , inhibits mutant-induced P29323 REA activity in 293T cells and growth of mutant-bearing Ba / P13726 REA cells . We also screened 85 NSCLC cell lines for Q02750 REA exon 2 mutations ; one line ( NCI-H 1437 ) harbors a Q56P substitution , a known transformation-competent allele of Q02750 REA originally identified in rat fibroblasts , and is sensitive to treatment with AZD 6244 . Q02750 REA mutants have not previously been reported in lung cancer and may provide a target for effective therapy in a small subset of patients with lung adenocarcinoma .

Procyanidin induces apoptosis of esophageal adenocarcinoma cells via JNK activation of c-Jun . Procyanidins are polymeric flavanols found in fruits and vegetables and have shown anticarcinogenic / chemopreventive properties . We previously showed that oligomeric procyanidin extracted from apples induced cell cycle arrest and apoptosis in esophageal adenocarcinoma ( OA ) cells . To understand the mechanism of action , we determined transcriptomic changes induced by procyanidin in OA cells . Pathway analysis implicated mitogen-activated protein kinase signaling pathways in eliciting these responses . Procyanidin induced the activation of JNK and p38 and the phosphorylation and expression of c-Jun . Inhibition of JNK but not p38 kinase activity prevented the procyanidin-induced phosphorylation and expression of c-Jun . Knockdown of the expression of P45983 REA , P45984 REA , or P05412 REA diminished procyanidin-induced effects on cell proliferation and apoptosis . c-Jun is a component of the transcription factor AP - 1 and AP - 1 binding sites are overrepresented in the promoters of procyanidin-induced genes . This indicates that JNK activation of c-Jun by procyanidin leads to the induction of apoptosis of OA cells and suggests a role for a c-Jun-mediated transcriptional program . These data provide a mechanistic understanding of how procyanidin specifically targets a distinct pathway involved in the induction of apoptosis in OA cells and will inform future studies investigating its use as a chemopreventive / therapeutic agent .

P10721 REA - D8 16 mutations in Q01196 REA - Q06455 - positive AML are associated with impaired event-free and overall survival . Mutations in codon D8 16 of the P10721 REA gene represent a recurrent genetic alteration in acute myeloid leukemia ( AML ) . To clarify the biologic implication of activation loop mutations of the P10721 REA gene , 1940 randomly selected AML patients were analyzed . In total , 33 ( 1.7 % ) of 1940 patients were positive for D8 16 mutations . Of these 33 patients , 8 ( 24.2 % ) had a t (8 ; 21 ) , which was significantly higher compared with the subgroup without D8 16 mutations . Analyses of genetic subgroups showed that P10721 REA - D8 16 mutations were associated with t (8 ; 21 ) / Q01196 REA - Q06455 and other rare Q01196 REA translocations . In contrast , other activating mutations like P36888 REA and P01111 REA mutations were very rarely detected in Q01196 REA - rearranged leukemia . P10721 REA mutations had an independent negative impact on overall ( median 304 vs 1836 days ; P = . 006 ) and event-free survival ( median 244 vs 744 days ; P = . 003 ) in patients with t (8 ; 21 ) but not in patients with a normal karyotype . The P10721 REA - D8 16V receptor expressed in Ba / P13726 REA cells was resistant to growth inhibition by the selective PTK inhibitors imatinib and SU5614 but fully sensitive to PKC 412 . Our findings clearly indicate that activating mutations of receptor tyrosine kinases are associated with distinct genetic subtypes in AML . The P10721 REA - D8 16 mutations confer a poor prognosis to Q01196 REA - Q06455 - positive AML and should therefore be included in the diagnostic workup . Patients with P10721 REA - D8 16 - positive / Q01196 REA - Q06455 - positive AML might benefit from early intensification of treatment or combination of conventional chemotherapy with P10721 REA PTK inhibitors .

DB01016 MEN exerts an antitumor activity through reactive oxygen species-c-jun NH2 - terminal kinase pathway in human gastric cancer cell line MGC - 803 . DB01016 MEN , a blocker of DB00171 - sensitive potassium ( K ( DB00171 ) ) channels , can suppress progression of many cancers , but the involved mechanism is unclear . Herein we reported that MGC - 803 cells expressed the K ( DB00171 ) channels composed of Kir 6.2 and Q09428 REA subunits . DB01016 MEN induced cellular viability decline , coupled with cell apoptosis and reactive oxygen species ( ROS ) generation in MGC - 803 cells . Meanwhile , glibenclamide increased NADPH oxidase catalytic subunit gp91 ( phox ) expression and superoxide anion ( O2 - ) generation , and caused mitochondrial respiration dysfunction in MGC - 803 cells , suggesting that glibenclamide induced an increase of ROS derived from NADPH oxidase and mitochondria . DB01016 MEN could also lead to loss of mitochondrial membrane potential , release of cytochrome c and apoptosis-inducing factor ( O95831 ) , and activation of c-jun NH2 - terminal kinase ( JNK ) in MGC - 803 cells . Pretreatment with antioxidant N-acetyl-l-cysteine ( Q9C000 REA ) prevented glibenclamide-induced JNK activation , apoptosis and cellular viability decline . Furthermore , glibenclamide greatly decreased the cellular viability , induced apoptosis and inhibited Akt activation in wild-type mouse embryonic fibroblast ( MEF ) cells but not in P45983 REA - / - or P45984 REA - / - MEF cells . Taken together , our study reveals that glibenclamide exerts an antitumor activity in MGC - 803 cells by activating ROS-dependent , JNK-driven cell apoptosis . These findings provide insights into the use of glibenclamide in the treatment of human gastric cancer .

P07332 REA kinase promotes mast cell recruitment to mammary tumors via the stem cell factor / P10721 REA receptor signaling axis . P10721 REA receptor is required for mast cell development , survival , and migration toward its ligand stem cell factor ( P21583 REA ) . Many solid tumors express P21583 REA and this leads to mast cell recruitment to tumors and release of mediators linked to tumor angiogenesis , growth , and metastasis . Here , we investigate whether P07332 REA protein-tyrosine kinase , a downstream effector of P10721 REA signaling in mast cells , is required for migration of mast cells toward P21583 REA - expressing mammary tumors . Using a novel agarose drop assay for chemotaxis of bone marrow-derived mast cells ( BMMC ) toward P21583 REA , we found that defects in chemotaxis of fes-null BMMCs correlated with disorganized microtubule networks in polarized cells . P07332 REA displayed partial colocalization with microtubules in polarized BMMCs and has at least two direct microtubule binding sites within its N-terminal F-BAR and SH2 domains . An oligomerization-disrupting mutation within the Fer / Q15642 REA homology-Bin / P49418 REA / Rvs ( F-BAR ) domain had no effect on microtubule binding , whereas microtubule binding to the SH2 domain was dependent on the phosphotyrosine-binding pocket . P07332 REA involvement in mast cell recruitment to tumors was tested using the AC2M2 mouse mammary carcinoma model . These tumor cells expressed P21583 REA and promoted BMMC recruitment in a P10721 REA - and P07332 REA - dependent manner . Engraftment of AC2M2 orthotopic and subcutaneous tumors in control or fes-null mice , revealed a key role for P07332 REA in recruitment of mast cells to the tumor periphery . This may contribute to the reduced tumor growth and metastases observed in fes-null mice compared with control mice . Taken together , P07332 REA is a potential therapeutic target to limit the progression of tumors with stromal mast cell involvement .

Transforming growth factor alpha-induced expression of type 1 plasminogen activator inhibitor in astrocytes rescues neurons from excitotoxicity . Although transforming growth factor ( TGF ) - alpha , a member of the epidermal growth factor ( P01133 REA ) family , has been shown to protect neurons against excitotoxic and ischemic brain injuries , its mechanism of action remains unknown . In the present study , we used in vitro models of apoptotic or necrotic paradigms demonstrating that TGF-alpha rescues neurons from N-methyl-D-aspartate ( DB01221 ) - induced excitotoxic death , with the obligatory presence of astrocytes . Because neuronal tissue-type plasminogen activator ( t-PA ) release was shown to potentiate DB01221 - induced excitotoxicity , we observed that TGF-alpha treatment reduced DB01221 - induced increase of t-PA activity in mixed cultures of neurons and astrocytes . In addition , we showed that although TGF-alpha induces activation of the extracellular signal-regulated kinases ( ERKs ) in astrocytes , it failed to activate Q8NFH3 / Q8TCB0 in neurons . Finally , we showed that TGF-alpha , by an P29323 REA - dependent mechanism , stimulates the astrocytic expression of P05121 REA , a t-PA inhibitor , which mediates the neuroprotective activity of TGF-alpha against DB01221 - mediated excitotoxic neuronal death . Taken together , we indicate that TGF-alpha rescues neurons from DB01221 - induced excitotoxicity in mixed cultures through inhibition of t-PA activity , involving P05121 REA overexpression by an P29323 REA - dependent pathway in astrocytes .

[ Angiogenesis and renal cell carcinoma ] . Developments in the knowledge of molecular biology of renal cell carcinoma ( RCC ) over the past 20 years have been identified . Angiogenesis is playing a key role in the physiopathology of RCC . Von Hippel-Lindau ( P40337 REA ) alterations , HIFalpha accumulation and vascular endothelial growth factor ( P15692 REA ) overexpression are important mediators of this process . Several stategies have been developped to target angiogenesis for the treatment of metastatic RCC . These include inhibition of P15692 REA receptors ( inhibition of the tyrosine kinase activity ) or binding to the P15692 REA protein . Several additional kinases inhibitions including PDGF receptors are also targeted . DB01268 ( SU11248 ) is an orally biovailable small molecule that has demonstrated superiority over interferon-alpha for the treatment of metastatic RCC . In a recent randomized phase III study conducted in 750 patients , the response rate to sunitinib was 31 % and to interferon 6 % . The median of progression free survival ( PFS ) was 11 months for sunitinib and 5 months for interferon ( p < 0.001 ) . DB00398 SUB ( BAY 43-9006 ) was found to inhibit Raf 1 , but also P35968 REA and 3 , Flt 3 , P09619 REA - a and b and c-kit , has been tested in a phase III study against placebo after one prior systemic therapy . The median of the time to progression ( TTP ) for sorafenib was 24 weeks versus 12 weeks for patients in the placebo arm ( p = 0,01 ) . Other molecules tested in metastatic RCC will be presented including axitinib , pazopanib and bevacizumab .

Cbl-b is a negative regulator of inflammatory cytokines produced by IgE-activated mast cells . c-Cbl and Cbl-b E3 ubiquitin ligases are abundantly expressed in hemopoietic cells where they negatively regulate the activity and levels of many cell surface receptors and associated signaling molecules . By comparing bone marrow-derived mast cells from c-Cbl and Cbl-b-deficient mice it has recently been shown that Cbl-b is the dominant family member for negatively regulating signaling responses from high-affinity IgE receptors . In this study , we suggest that a possible reason for the greater enhancement of IgE receptor signaling in Cbl-b-deficient mice is the relatively higher levels of Cbl-b protein over c-Cbl in mast cells compared with other hemopoietic cells . We also directly compare mast cells from c-Cbl and Cbl-b-deficient mice and find that loss of Cbl-b , but not c-Cbl , increases cell growth , retards receptor internalization , and causes the sustained tyrosine phosphorylation of Syk and its substrates . However , loss of Cbl-b does not enhance the activation of P29323 REA or Akt , nor does it promote a greater calcium response . Furthermore , loss of Cbl-b or c-Cbl does not increase levels of the Syk or Lyn protein tyrosine kinases . Most notable , however , is the extremely large increase in the production of proinflammatory cytokines P01375 REA , P05231 REA , and P13500 REA by Cbl-b ( - / - ) mast cells compared with levels produced by c-Cbl ( - / - ) or wild-type cells . This marked induction , which appears to be restricted to these three cytokines , is dependent on IgE receptor activation and correlates with enhanced O15111 REA phosphorylation . Thus , Cbl-b functions as a potent negative regulator of cytokines that promote allergic and inflammatory reactions .

Suppression of P05121 REA expression through inhibition of the P00533 REA - mediated signaling cascade in rat kidney fibroblast by ascofuranone . Fibrosis in glomerulosclerosis causes progressive loss of renal function . Transforming growth factor ( TGF ) - beta , one of the major profibrotic cytokines , induces the synthesis of plasminogen activator inhibitor ( P05121 REA ) - 1 , a factor that plays a crucial role in the development of fibrosis . Here , we found that an isoprenoid antibiotic , ascofuranone , suppresses expression of profibrotic factors including matrix proteins and P05121 REA induced by TGF-beta in renal fibroblasts . Ascofuranone selectively inhibits phosphorylation of epidermal growth factor receptor ( P00533 REA ) , and downstream kinases such as P04049 REA , MEK -1/2 , and P27361 REA / 2 . P05121 REA transcription also is suppressed by treatment with kinase inhibitors for MEK -1/2 or P00533 REA , and with small interfering RNA for P00533 REA . Ascofuranone inhibits cellular metalloproteinase activity , and an inhibitor of metalloproteinases suppresses P00533 REA phosphorylation and P05121 REA transcription . These results suggest that ascofuranone suppresses expression of profibrotic factors through the inhibition of an P00533 REA - dependent signal transduction pathway activated by metalloproteinases .

DB00398 SUB as treatment for relapsed or refractory pediatric acute myelogenous leukemia . The prognosis for children with acute myelogenous leukemia ( AML ) has improved with overall survival rates of up to 65 % [ Pui et al . J Clin Oncol 2011 ; 29 : 551-565 ] . However , the cure rate for AML lags behind that of acute lymphoblastic leukemia . Advances in AML leukemogenesis are leading to refined risk stratification . P07333 REA like tyrosine kinase 3 ( P36888 REA ) mutations are independently associated with a poor prognosis . Newer kinase inhibitors , including sorafenib , have shown promise in adult studies . We report three pediatric patients with relapsed AML who achieved a sustained remission with sorafenib . Further trials are necessary to understand the role of sorafenib in pediatric AML .

DB00398 SUB does not improve efficacy of chemotherapy in advanced pancreatic cancer : A GISCAD randomized phase II study . BACKGROUND : The RAF-MEK - P29323 REA pathway is commonly activated in pancreatic cancer because of a high frequency of P01116 REA - P15056 REA mutations . A phase II randomized trial was designed to investigate the activity of sorafenib in combination with chemotherapy in advanced pancreatic cancer . METHODS : Locally advanced or metastatic pancreatic adenocarcinoma patients were randomized in a 1:1 ratio to receive cisplatin plus gemcitabine with sorafenib 400mg bid ( arm A ) or without sorafenib ( arm B ) . RESULTS : One hundred and fourteen patients were enrolled ; of these , 43 ( 74.6 % ) patients progressed in arm A and 44 ( 82.4 % ) in arm B . Median progression-free survival was 4.3 months ( 95 % CI : 2.7- 6.5 ) and 4.5 months ( 95 % CI : 2.5- 5.2 ) , respectively ( HR = 0.92 ; 95 % CI : 0.62- 1.35 ) . Median overall survival was 7.5 ( 95 % CI : 5.6- 9.7 ) and 8.3 months ( 95 % CI : 6.2- 8.7 ) , respectively ( HR = 0.95 ; 95 % CI : 0.62- 1.48 ) . Response rates were 3.4 % in arm A and 3.6 % in arm B . CONCLUSIONS : DB00398 SUB does not significantly enhance activity of chemotherapy in advanced pancreatic cancer patients , and therefore should not be assessed in phase III trials .

5 - Azacitidine restores and amplifies the bicalutamide response on preclinical models of androgen receptor expressing or deficient prostate tumors . BACKGROUND : Epigenetic modifications play a key role in the in prostate cancer ( Pca ) progression to a hormone refractory state ( HRPC ) and the current use of agents targeting epigenetic changes has become a topic of intense interest in cancer research . In this regard , 5 - Azacitine ( 5 - Aza ) represents a promising epigenetic modulator . This study tested the hypothesis that 5 - Aza may restore and enhance the responsiveness of HRPC cells to anti-hormonal therapy on P10275 REA ( AR ) expressing ( 22rv1 ) and AR-deficient ( PC3 ) cells . METHODS : The effects were studied in vitro and in vivo models . This sequential treatment induced in vitro cell cycle arrest and apoptosis both in 22rv1 and PC3 tumor cell lines . RESULTS : This combined treatment up-regulated the expression of P48023 REA , phospho - Q13158 REA , p16 ( INKA ) , Bax , Bak , and P38936 REA ( P38936 REA ) , and inhibited FLIP , Bcl - 2 , and Bcl-XL expression . The re-activation of hormonal response of AR-negative PC3 cell line was partially due to the AR re-expression mediated by 5 - Aza treatment . In contrast , the increase in the response to anti-androgenic therapy in 22rv1 did not correlate with AR expression levels . Furthermore , xenograft studies revealed that the combined treatment of 5 - Aza with AR-antagonist DB01128 MEN had additive / synergistic effects in repressing tumor growth in vivo and the underlying mechanisms responsible for these effects seem to be in part mediated by induction of apoptosis . CONCLUSIONS : So , this study strongly suggests a therapeutic potential of 5 - Aza in combination with anti-androgen therapy in patients with in AR expressing and AR-deficient HRPC .

[ DB00398 SUB in combination with chemotherapy in the induction therapy for P36888 REA - ITD positive acute monocytic leukemia : a case report and literature review ] . OBJECTIVE : To explore the safety and efficacy of sorafenib in combination with chemotherapy for the treatment of P36888 REA positive acute myeloid leukemia ( AML ) , to highlight the impact of P36888 REA mutations and targeting therapy on response of AML . METHODS : The clinical and laboratory features and the treatment response , especially the safety profile of sorafenib in an acute monocytic leukemia patient with P17948 REA - ITD were reported . RESULTS : The patient achieved clinical and molecular CR after sorafenib was added to the second course of combination chemotherapy . The side effects of sorafenib were mild and tolerable . CONCLUSION : The patient responded well to the combination of sorafenib and standard chemotherapy of AML without significant adverse effects .

Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 - R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 REA ) in the presence of cycloheximide ( Reid , T . R . , Torti , F . , and Ringold , G . M . ( 1989 ) J . Biol . Chem . 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 REA or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 REA - resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6 - desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 - R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 REA in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 REA and is essential to the rapid cytotoxic response elicited by P01375 REA in the absence of protein synthesis in wild-type Q96RJ0 cells .

Multiple mechanisms mediate resistance to sorafenib in urothelial cancer . Genetic and epigenetic changes in the mitogen activated protein kinase ( MAPK ) signaling render urothelial cancer a potential target for tyrosine kinase inhibitor ( TKI ) treatment . However , clinical trials of several TKIs failed to prove efficacy . In this context , we investigated changes in MAPK signaling activity , downstream apoptotic regulators and changes in cell cycle distribution in different urothelial cancer cell lines ( UCCs ) upon treatment with the multikinase inhibitor sorafenib . None of the classical sorafenib targets ( vascular endothelial growth factor receptor 1 / - receptor 2 , P17948 REA / - R2 ; platelet-derived growth factor receptor α / - receptor β , P09619 REA - α / - β ; c - P10721 REA ) was expressed at significant levels leaving RAF proteins as its likely molecular target . Low sorafenib concentrations paradoxically increased cell viability , whereas higher concentrations induced P55008 arrest and eventually apoptosis . MAPK signaling remained partly active after sorafenib treatment , especially in T24 cells with an oncogenic P01112 REA mutation . AKT phosphorylation was increased , suggesting compensatory activation of the phosphatidylinositol - 3 - kinase ( PI3K ) pathway . DB00398 SUB regularly down regulated the anti-apoptotic myeloid cell leukemia 1 ( Mcl - 1 ) protein , but combinatorial treatment with ABT - 737 targeting other B-cell lymphoma 2 ( Bcl - 2 ) family proteins did not result in synergistic effects . In summary , efficacy of sorafenib in urothelial cancer cell lines appears hampered by limited effects on MAPK signaling , crosstalk with further cancer pathways and an anti-apoptotic state of UCCs . These observations may account for the lack of efficacy of sorafenib in clinical trials and should be considered more broadly in the development of signaling pathway inhibitors for drug therapy in urothelial carcinoma .

Regulation of gene expression in melanoma : new approaches for treatment . The molecular changes associated with the transition of melanoma cells from radial growth phase ( RGP ) to vertical growth phase ( VGP , metastatic phenotype ) are not yet well defined . We have demonstrated that the progression of human melanoma is associated with loss of expression of the transcription factor P05549 REA . In metastatic melanoma cells , this loss resulted in overexpression of P43121 / P43121 , P08253 REA , the thrombin receptor ( P25116 REA ) , and lack of c - P10721 REA expression . The transition from RGP to VGP is also associated with overexpression of the angiogenic factor P10145 REA . Additionally , the transition of melanoma cells from RGP to VGP is associated with overexpression of the transcription factors CREB and P39905 REA - 1 , both of which may act as survival factors for human melanoma cells . Inactivation of CREB / P39905 REA - 1 activities in metastatic melanoma cells by dominant-negative CREB or by anti - P39905 REA - 1 single chain antibody fragment ( ScFv ) , resulted in deregulation of P08253 REA and P43121 / P43121 , increased the sensitivity of melanoma cells to apoptosis , and inhibition of their tumorigenicity and metastatic potential in vivo . In this prospect article , we summarize our data on the role of P05549 REA and CREB / P39905 REA - 1 in the progression of human melanoma and report on the development of new fully human antibodies anti - P43121 / P43121 and anti - P10145 REA which could serve as new modalities for the treatment of melanoma .

DB00398 SUB : targeting multiple tyrosine kinases in cancer . DB00398 SUB ( BAY 43-9006 , Nexavar ® ) is an oral multiple tyrosine kinase inhibitor . Main targets are receptor tyrosine kinase pathways frequently deregulated in cancer such as the Raf-Ras pathway , vascular endothelial growth factor ( P15692 REA ) pathway , and P07333 REA - like tyrosine kinase 3 ( P36888 REA ) . DB00398 SUB was approved by the FDA in fast track for advanced renal cell cancer and hepatocellular cancer and shows good clinical activity in thyroid cancer . Multiple clinical trials are undertaken to further investigate the role of sorafenib alone or in combination for the treatment of various tumor entities .

DB09301 glycosaminoglycans as major P16109 REA ligands on metastatic breast cancer cell lines . The metastatic breast cancer cell line , 4T1 , abundantly expresses the oligosaccharide sialylated Lewis x ( sLe ( x ) ) . SLe ( x ) oligosaccharide on tumor cells can be recognized by E - and P16109 REA , contributing to tumor metastatic process . We observed that both selectins reacted with this cell line . However , contrary to the P16581 REA reactivity , which was sLe ( x ) dependent , P16109 REA reactivity with this cell line was sLe ( x ) - independent . The sLe ( x ) - Neg variant of the 4T1 cell line with markedly diminished expression of sLe ( x ) and lack of sLe ( a ) , provided a unique opportunity to characterize P16109 REA ligands and their contribution to metastasis in the absence of overlapping selectin ligands and P16581 REA binding . We observed that P16109 REA binding was Ca ( 2 + ) - independent and sulfation-dependent . We found that P16109 REA reacted primarily with cell surface chondroitin sulfate ( CS ) proteoglycans , which were abundantly and stably expressed on the surface of the 4T1 cell line . P16109 REA binding to the 4T1 cells was inhibited by heparin and CS glycosaminoglycans ( GAGs ) . Moreover , DB01109 MEN administration significantly inhibited experimental lung metastasis . In addition , the data suggest that surface CS GAG chains were involved in P16109 REA mediated adhesion of the 4T1 cells to murine platelets and human umbilical vein endothelial cells . The data suggest that CS GAGs are also the major P16109 REA - reactive ligands on the surface of human MDA-MET cells . The results warrant conducting clinical studies on the involvement of cell surface CS chains in breast cancer metastasis and evaluation of various CS types and their biosynthetic pathways as target for development of treatment strategies for antimetastatic therapy of this disease .

Peripheral medulloepithelioma : a rare tumor with a potential target therapy . BACKGROUND : Medulloepithelioma ( ME ) is a rare embryonal tumor predominantly located in the eye or in the central nervous system without an established treatment . CASE PRESENTATION : We report of a case of a localized peripheral ME treated with conventional and high dose chemotherapy , surgery and local radiotherapy . At relapse , the tumor tissue revealed a different molecular signature compared to the initial tumor mass . This molecular signature revealed a high expression of platelet derived growth factor receptor ( P09619 REA ) . DB00398 SUB plus irinotecan and temozolomide was started with a 5 month progression free survival . CONCLUSION : Our experience suggests a possible role of sorafenib or different P09619 REA inhibitors in ME . Targeting treatment could represent an adjuvant and / or alternative therapy for ME and other rare tumors .

Iodide - and glucose-handling gene expression regulated by sorafenib or cabozantinib in papillary thyroid cancer . CONTEXT : The aberrant silencing of iodide-handling genes accompanied by up-regulation of glucose metabolism presents a major challenge for radioiodine treatment of papillary thyroid cancer ( PTC ) . OBJECTIVE : This study aimed to evaluate the effect of tyrosine kinase inhibitors on iodide-handling and glucose-handling gene expression in BHP 2-7 cells harboring P07949 REA / Q13635 REA rearrangement . MAIN OUTCOME MEASURES : In this in vitro study , the effects of sorafenib or cabozantinib on cell growth , cycles , and apoptosis were investigated by cell proliferation assay , cell cycle analysis , and P08758 REA - FITC apoptosis assay , respectively . The effect of both agents on signal transduction pathways was evaluated using the Western blot . Quantitative real-time PCR , Western blot , immunofluorescence , and radioisotope uptake assays were used to assess iodide-handling and glucose-handling gene expression . RESULTS : Both compounds inhibited cell proliferation in a time-dependent and dose-dependent manner and caused cell cycle arrest in the G0 / P55008 phase . DB00398 SUB blocked P07949 REA , AKT , and P27361 REA / 2 phosphorylation , whereas cabozantinib blocked P07949 REA and AKT phosphorylation . The restoration of iodide-handling gene expression and inhibition of glucose transporter 1 and 3 expression could be induced by either drug . The robust expression of sodium / iodide symporter induced by either agent was confirmed , and ( 125 ) I uptake was correspondingly enhanced . ( 18 ) F - DB09150 accumulation was significantly decreased after treatment by either sorafenib or cabozantinib . CONCLUSIONS : DB00398 SUB and cabozantinib had marked effects on cell proliferation , cell cycle arrest , and signal transduction pathways in PTC cells harboring P07949 REA / Q13635 REA rearrangement . Both agents could be potentially used to enhance the expression of iodide-handling genes and inhibit the expression of glucose transporter genes .

Suppression of androgen receptor-mediated gene expression by a sequence-specific DNA-binding polyamide . P10275 REA ( AR ) is essential for the growth and progression of prostate cancer in both hormone-sensitive and hormone-refractory disease . A DNA-binding polyamide that targets the consensus androgen response element binds the prostate-specific antigen ( PSA ) promoter androgen response element , inhibits androgen-induced expression of PSA and several other AR-regulated genes in cultured prostate cancer cells , and reduces AR occupancy at the PSA promoter and enhancer . Down-regulation of PSA by this polyamide was comparable to that produced by the synthetic antiandrogen bicalutamide ( DB01128 MEN ) at the same concentration . Genome-wide expression analysis reveals that a similar number of transcripts are affected by treatment with the polyamide and with bicalutamide . Direct inhibition of the AR-DNA interface by sequence-specific DNA binding small molecules could offer an alternative approach to antagonizing AR activity .

DB00398 SUB in patients with metastatic gastrointestinal stromal tumors who failed two or more prior tyrosine kinase inhibitors : a phase II study of Korean gastrointestinal stromal tumors study group . PURPOSE : To evaluated the efficacy and safety of sorafenib in patients with advanced gastrointestinal stromal tumors ( GIST ) who failed to previous standard treatments . EXPERIMENTAL DESIGN : Thirty-one patients with measurable metastatic GIST who failed both imatinib and sunitinib were accrued . DB00398 SUB was administered orally at 400 mg twice daily until disease progression or development of intolerance . The primary endpoint was disease control rate ( response + stable disease , DCR ) at 24 weeks . RESULTS : DB00398 SUB was well tolerated , with hand-foot skin reaction , fatigue , hypertension , and abdominal pain being the most frequent adverse events . The relative dose intensity of sorafenib during the first 6 months was > 80 % . Four patients achieved partial response ( response rate 13 % , 95 % CI 1-25 % ) , and 16 ( 52 % ) had stable disease . DCR at 24 weeks was measured as 36 % ( 95 % CI 19-52 % ) . Median progression-free and overall survivals were 4.9 and 9.7 months , respectively . Progression-free survival of patients with prior use of nilotinib ( P = . 0085 ) and with primary genotypes other than P10721 REA exon 11 mutation ( P = . 0341 ) was significantly shorter than that of patients without . CONCLUSIONS : DB00398 SUB showed antitumor activity in this population of imatinib and sunitinib pretreated GIST . With sorafenib , about one third of patients can maintain disease control for more than 24 weeks .

DB00398 SUB for the treatment of hepatocellular carcinoma across geographic regions . DB00398 SUB is an oral multikinase inhibitor targeting Raf , P15692 REA receptor , PDGF receptor , c-kit , Flt - 3 and rearranged during transfection ( P07949 REA ) . Two randomized , placebo-controlled trials for Western and Asian patients , respectively , demonstrated that sorafenib significantly prolongs overall survival and time to progression in patients with advanced hepatocellular carcinoma ( HCC ) . These have become the reference treatment for future clinical trials of advanced HCC . DB00398 SUB is well tolerated in patients with Child-Pugh liver function class A , but limited data are available in Child-Pugh class B and C patients . Clinical trials are ongoing to test the efficacy of sorafenib-based combination therapy and sorafenib adjuvant therapy for HCC .

DB00398 SUB ( Nexavar ) induces molecular remission and regression of extramedullary disease in a patient with P36888 REA - ITD + acute myeloid leukemia . The fms-related tyrosine kinase 3 internal tandem duplication ( P36888 REA - ITD ) can be found in about one quarter of patients with acute myeloid leukemia ( AML ) [ Small D . P36888 REA mutations : biology and treatment . Hematology Am Soc Hematology . Educ . Program 2006 ; 178-84 [ Review ] ] . Patients who carry this mutation have a high risk of relapse even after allogeneic stem cell transplantation [ Sheikhha MH , Awan A , Tobal K , Liu Yin JA . Prognostic significance of P36888 REA ITD and D8 35 mutations in AML patients . Hematol J 2003 ; 4:41- 6 ; Meshinchi S , Arceci RJ , Sanders JE , Smith FO , Woods WB , Radich JP , et al . Role of allogeneic stem cell transplantation in P36888 REA / ITD-positive AML . Blood 2006 ; 108 ( 1 ): 400-1 ] . Recent reports show that DB00398 SUB , a multikinase inhibitor has significant activity against P36888 REA - ITD ( + ) blasts in vitro [ Auclair D , Miller D , Yatsula V , Pickett W , Carter C , Chang Y , et al . Antitumor activity of sorafenib in P36888 REA - driven leukemic cells . Leukemia 2007 ; 21 ( 3 ): 439-45 ] . We here report the first clinical case of molecular remission induced by DB00398 SUB in a patient with P36888 REA - ITD ( + ) AML and extramedullary disease after allogenic stem cell transplantation .

The ability of sorafenib to inhibit oncogenic PDGFRbeta and P36888 REA mutants and overcome resistance to other small molecule inhibitors . BACKGROUND AND OBJECTIVES : Activated tyrosine kinases are implicated in the pathogenesis of chronic and acute leukemia , and represent attractive targets for therapy . DB00398 SUB ( BAY 43-9006 , Nexavar ) is a small molecule B-RAF inhibitor that is used for the treatment of renal cell carcinoma , and has been shown to have activity against receptor tyrosine kinases from the platelet-derived growth factor receptor ( P09619 REA ) and vascular endothelial growth factor receptor ( VEGFR ) families . We investigated the efficacy of sorafenib at inhibiting mutants of the receptor tyrosine kinases PDGFRbeta , P10721 REA , and P36888 REA , which are implicated in the pathogenesis of myeloid malignancies . DESIGN AND METHODS : We tested the effect of sorafenib on the proliferation of hematopoietic cells transformed by P41212 REA - PDGFRbeta , P36888 REA with an internal tandem duplication or D8 35Y point mutation , and the P10721 REA ( D8 16V ) mutant . The direct effect of sorafenib on the activity of these kinases and their downstream signaling was tested using phospho-specific antibodies . RESULTS : We show that sorafenib is a potent inhibitor of P41212 REA - PDGFRbeta and P36888 REA mutants , including some of the mutants that confer resistance to PKC 412 and other P36888 REA inhibitors . DB00398 SUB induced a cell cycle block and apoptosis in the acute myeloid leukemia cell lines MV4 - 11 and MOLM - 13 , both expressing P36888 REA with an internal tandem duplication , whereas no effect was observed on four other acute myeloid leukemia cell lines . The imatinib-resistant P10721 REA ( D8 16V ) mutant , associated with systemic mastocytosis , was found to be resistant to sorafenib . INTERPRETATION AND CONCLUSIONS : These results warrant further clinical studies of sorafenib for the treatment of myeloid malignancies expressing activated forms of PDGFRbeta and P36888 REA .

Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 MEN or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling .

[ New therapies targeting the genetic mutations responsible for different types of melanoma ] . A number of molecular alterations have been described for melanoma . Melanomas with P15056 REA mutations tend to be located in areas of intermittent sun exposure , whereas melanomas with P10721 REA mutations mostly appear in acral areas , the mucosas , and areas of chronic sun exposure . DB00398 SUB , a P15056 REA inhibitor , has a cytostatic effect on most melanomas with mutations affecting the mitogen-activated protein kinase ( MAPK ) pathway , and is also capable of triggering apoptosis in a small subgroup of these melanomas . By inhibiting P10721 REA , imatinib has a cytostatic and cytotoxic effect on melanomas with P10721 REA mutations , and probably has the same effect on another subgroup of melanomas with other as yet imperfectly understood P10721 REA mutations . For therapy to be effective , agents should be selected according to the pathways associated with the genetic mutations present in the melanoma .

DB00398 SUB inhibits the angiogenesis and growth of orthotopic anaplastic thyroid carcinoma xenografts in nude mice . Anaplastic thyroid carcinoma ( ATC ) remains one of the most lethal human cancers . We hypothesized that sorafenib , a multikinase inhibitor of the BRaf , vascular endothelial growth factor receptor - 2 , and platelet-derived growth factor receptor-beta kinase , would decrease tumor growth and angiogenesis in an orthotopic model of ATC . The in vitro antiproliferative and proapoptotic effects of sorafenib on ATC cell lines were examined . To study the in vivo effects of sorafenib on orthotopic ATC tumors in nude mice , sorafenib was given p . o . at 40 or 80 mg / kg daily . Intratumoral effects were studied using immunohistochemical analysis . The effect of sorafenib on survival of the mice was also studied . DB00398 SUB inhibited the in vitro proliferation of ATC cell lines . DB00398 SUB also significantly inhibited tumor angiogenesis via the induction of endothelial apoptosis in an orthotopic model of thyroid cancer . As result , the growth of orthotopic ATC xenografts was reduced and the survival of the test animals was improved . DB00398 SUB exerts significant antitumor activity in an orthotopic xenograft model of ATC via a potent antiangiogenic effect . The antiangiogenic effects of sorafenib suggest that its use in clinical setting may not depend on the P15056 REA mutational status of thyroid tumors . Given the lack of curative options for patients with ATC , sorafenib warrants further study as a therapeutic agent against ATC .

DB00107 alleviates the neuroendocrine and cytokine response to bacterial endotoxin in healthy men . DB00107 is a hormone and neurotransmitter found to have anti-inflammatory functions in rodents . Here we used experimental bacterial endotoxinemia to examine the role of exogenous oxytocin administration on innate immune responses in humans . Ten healthy men received , in a randomized , placebo-controlled , crossover design , placebo , oxytocin , LPS , and LPS + oxytocin . DB00107 treatment resulted in a transient or prolonged reduction of endotoxin-induced increases in plasma DB01285 , cortisol , procalcitonin , P01375 REA , IL - 1 receptor antagonist , P05112 REA , P05231 REA , macrophage inflammatory protein - 1alpha , macrophage inflammatory protein - 1beta , monocyte chemoattractant protein - 1 ( P13500 REA ) , interferon-inducible protein 10 , and P15692 REA . In vitro , oxytocin had no impact on LPS effects in releasing P01375 REA , P05231 REA , and P13500 REA in monocytes and peripheral blood mononuclear cells from healthy human donors . In summary , oxytocin decreases the neuroendocrine and cytokine activation caused by bacterial endotoxin in men , possibly due to the pharmacological modulation of the cholinergic anti-inflammatory pathway . DB00107 might be a candidate for the therapy of inflammatory diseases and conditions associated with high cytokine and P15692 REA levels .

Functional and molecular characterization of ex vivo cultured epiretinal membrane cells from human proliferative diabetic retinopathy . Characterization of the cell surface marker phenotype of ex vivo cultured cells growing out of human fibrovascular epiretinal membranes ( fvERMs ) from proliferative diabetic retinopathy ( PDR ) can give insight into their function in immunity , angiogenesis , and retinal detachment . FvERMs from uneventful vitrectomies due to PDR were cultured adherently ex vivo . Surface marker analysis , release of immunity - and angiogenesis-pathway-related factors upon P01375 REA α activation and measurement of the intracellular calcium dynamics upon mechano-stimulation using fluorescent dye Fura - 2 were all performed . FvERMs formed proliferating cell monolayers when cultured ex vivo , which were negative for endothelial cell markers ( CD31 , P35968 REA ) , partially positive for hematopoietic - ( P28906 REA , Q08722 REA ) and mesenchymal stem cell markers ( CD73 , CD90 / Thy - 1 , and P09619 REA β ) , and negative for CD105 . CD146 / P43121 and CD166 / Q13740 REA , previously unreported in cells from fvERMs , were also expressed . Secretion of 11 angiogenesis-related factors ( DPPIV / P27487 REA , P58294 REA / P30613 REA , ET - 1 , P18065 REA and 3 , P10145 REA / P10145 REA , P13500 REA / P13500 REA , P14780 REA , PTX 3 / P26022 REA , P05121 REA / P05121 REA , P36955 / P36955 , P01033 REA , and P07996 REA - 1 ) were detected upon P01375 REA α activation of fvERM cells . Mechano-stimulation of these cells induced intracellular calcium propagation representing functional viability and role of these cells in tractional retinal detachment , thus serving as a model for studying tractional forces present in fvERMs in PDR ex vivo .

DB06643 MEN for joints and bones . DB06643 MEN is an investigational , fully human monoclonal antibody with a high affinity and specificity for receptor activator of nuclear factor kappaB ligand ( O14788 REA ) , a cytokine member of the tumor necrosis factor family . O14788 REA , an essential mediator of osteoclast formation , function , and survival , plays a major role in the pathogenesis of postmenopausal osteoporosis , structural damage in rheumatoid arthritis , and bone loss associated with other skeletal disorders . DB06643 MEN suppresses bone turnover by inhibiting the action of O14788 REA on osteoclasts . DB06643 MENMAX DB06643 MEN reduces bone turnover and increases bone mineral density in postmenopausal women with low bone mineral density , reduces fracture risk in women with postmenopausal osteoporosis , and inhibits structural damage in patients with rheumatoid arthritis when added to ongoing methotrexate treatment . It is generally well tolerated , with a good safety profile . Adverse and serious adverse events , including infections and malignancy , are similar in patients treated with denosumab or placebo .

DB00398 SUB - associated remission of psoriasis in hypernephroma : case report . Psoriasis is a disease characterized by epidermal hyperproliferation that results in the formation of lesional plaques covered by scale . Psoriasis is thought to be angiogenesis dependent . Clear cell renal cell carcinoma is a hypervascularized solid tumor associated with loss of function of the von Hippel-Lindau ( P40337 REA ) tumor suppressor gene and increased P04049 REA activity . A 68 - year-old man who suffered from recalcitrant psoriasis for over 50 years was treated with sorafenib for metastatic clear cell renal carcinoma . One month later , his psoriasis , previously 8 x 6 cm on the mid posterior thorax , completely resolved . DB00398 SUB works by inhibiting several receptor tyrosine kinases ( RTKs ) , such as vascular endothelial growth factor ( VEGFR ) and platelet-derived growth factor receptor ( P09619 REA ) ) . It also inhibits intracellular Raf kinase ( P04049 REA ) , which targets the ubiquitous mitogen-activated protein kinase ( MAPK ) intracellular signal transduction pathway . We suggest that this patient ' s remission of psoriasis could be related to the inhibition / modulation of P15692 REA , P09619 REA , P04049 REA , and MAPK .

DB00398 SUB tosylate as a radiosensitizer in malignant astrocytoma . Progress in research on the molecular aspects of glioblastoma has yet to provide a medical therapy that significantly improves prognosis . Glioblastoma invariably progress through current treatment regimens with radiotherapy as a key component . Activation of several signaling pathways is thought to be associated with this resistance to radiotherapy . Ras activity is exceptionally high in glioblastoma and may regulate sensitivity to radiotherapy . P04049 REA , a downstream effector of Ras , demonstrates a high amount of activity in glioblastoma . Therefore , P04049 REA inhibition should be considered as a mechanism to increase the effectiveness of radiotherapy in treatment regimen . In vitro analysis was performed with a novel P04049 REA kinase inhibitor ( BAY 54-9085 ) in culture with the glioblastoma cell line U1242 . The cell line was treated in serum-containing media and analyzed for the effect of the BAY 54-9085 alone and BAY 54-9085 combined with radiation on cell death . BAY 54-9085 displayed a cytocidal effect on glioblastoma cells following a 3 day incubation with the drug in serum-containing media . A dose of 2.5 μM displayed moderate cell death which significantly increased with a dose of 5.0 μM . In addition , glioblastoma cells treated with both the BAY 54-9085 and gamma radiation displayed a significant increase in cell death ( 85.5 % ) as compared to either BAY 54-9085 ( 73.1 % ) or radiation ( 34.4 % ) alone . Radiation therapy is a key component of treatment for glioblastoma . A novel P04049 REA inhibitor displayed in vitro evidence of synergistically increasing cell death of glioblastoma cells in combination with radiation .

Prasugrel : a new antiplatelet drug for the prevention and treatment of cardiovascular disease . Prasugrel , trade name DB06209 MEN , is an investigational new antiplatelet drug currently under review for clinical use by the Food and Drug Administration . It is a thienopyridine analog with a structure similar to that of clopidogrel and ticlopidine . Thienopyridine derivatives inhibit platelet aggregation induced by adenosine diphosphate by irreversibly inhibiting the binding of adenosine diphosphate to the purinergic Q9H244 REA receptor on the platelet surface . Prasugrel has been shown to be a potent antiplatelet agent with a faster , more consistent , and greater inhibition of platelet aggregation compared with clopidogrel . It is debatable , however , how effectively these pharmacologic benefits will translate to clinical benefits . The results of the large TRITON-TIMI 38 trial , which compared prasugrel and clopidogrel in patients with acute coronary syndrome who were scheduled to receive coronary stents , demonstrated a significant reduction in ischemic events , including stent thrombosis , with prasugrel , but with an increased risk of major bleeding . The exact role of prasugrel in the management of ischemic heart disease is still being defined , but the risk : benefit ratio will likely play a major role in directing the best place for therapy with this new agent .

Antifibrotic activity of sorafenib in experimental hepatic fibrosis : refinement of inhibitory targets , dosing , and window of efficacy in vivo . BACKGROUND : DB00398 SUB , which is approved for treatment of HCC , has also shown promising antifibrotic activity , and therefore refinement of its dosing requirements and window of efficacy are important goals prior to antifibrotic clinical trials . AIM : The purpose of this study was to determine the minimal effective dose and optimal timing of sorafenib therapy in cultured human stellate cells and in rats with experimental hepatic fibrosis . METHODS : Effects of sorafenib were assessed in a human stellate cell line ( LX - 2 ) . In vivo , rats were treated for 8 weeks with TAA three times per week ( 150 mg / kg IP ) , and with either PBS or sorafenib administered daily at doses of 1.25 , 5 or 7 mg / kg / day gavage either at the beginning of TAA administration for 8 weeks , during weeks 4-8 , or from weeks 8-12 . RESULTS : DB00398 SUB treatment significantly inhibited LX - 2 proliferation by > 75 % ( 7.5 or 15 μM ) . Treatment with 7.5- μM sorafenib for 12 h markedly inhibited expression of TGFβ 1 , P01033 REA , collagen I , and P08253 REA mRNAs , but not of β - P09619 REA or type I TGFβR . In vivo , sorafenib significantly inhibited liver fibrosis when started concurrently with TAA and during weeks 4-8 with TAA . In contrast , there was no significant effect of sorafenib on fibrogenic gene expression or fibrosis when begun after cirrhosis was already established . CONCLUSION : DB00398 SUB is anti-proliferative and antifibrotic towards human HSCs in culture , and is a potent antifibrotic agent in TAA-induced hepatic fibrosis in rats . The drug is effective at relatively low doses at the early stage of liver fibrosis , but is not effective when cirrhosis is already established .

P25116 REA genotype influences platelet aggregation and procoagulant responses in patients with coronary artery disease prior to and during clopidogrel therapy . Genetic variations of the protease-activated receptor - 1 ( P25116 REA ) have been associated with platelet receptor density and linked to thrombin receptor-activating peptide ( TRAP ) - induced phenotypes of platelet aggregation and P16109 REA expression . We investigated whether the P25116 REA intervening sequence - 14 A > T dimorphism influences platelet procoagulant activity . We also determined whether the Q9H244 REA antagonist clopidogrel could offset any observed functional polymorphism of the P25116 REA receptor by inhibiting Q9H244 REA - mediated amplification of TRAP-induced responses . We studied 54 patients listed for elective percutaneous coronary intervention assessing TRAP-induced platelet aggregation and markers of procoagulant activity . Platelet responses were measured at baseline , 4 h post clopidogrel 300 mg , and 10 and 28 days following clopidogrel 75 mg daily . Each patient was genotyped for the P25116 REA intervening sequence - 14 A / T dimorphism . Increased platelet aggregation and procoagulant responses were observed with P25116 REA A allele homozygotes . DB00758 significantly inhibited these platelet responses regardless of P25116 REA genotype , but did not offset the hyper-reactivity associated with the A / A homozygotes . We conclude that a common sequence variation within the P25116 REA gene influences TRAP-induced platelet procoagulant activity as well as aggregation . Higher platelet reactivity associated with P25116 REA IVSn - 14 A allele homozygotes persists despite clopidogrel therapy . These individuals may be at higher risk of thromboembolic events and may require additional anti-platelet medication .

P01375 REA - alpha blocks apoptosis in melanoma cells when P15056 REA signaling is inhibited . The protein kinase P15056 REA , a component of the DB01367 / RAF / mitogen-activated protein kinase / extracellular signal-regulated kinase ( P29323 REA ) kinase ( MEK ) / P29323 REA signaling pathway , regulates cell fate in response to extracellular signals . Activating mutations in P15056 REA occur in approximately 70 % of human melanomas . The active proteins stimulate constitutive pathway signaling , proliferation , and survival . Thus , inhibition of P15056 REA signaling in melanoma cells causes cell cycle arrest and induces cell death through apoptosis , validating P15056 REA as an important therapeutic target . Here , we show that the apoptosis induced by inhibition of P15056 REA signaling in melanoma cells can be prevented if the cells are treated with tumor necrosis factor ( P01375 REA ) - alpha . This allows the cells to recover from the inhibition of P15056 REA signaling and reenter the cell cycle . This effect occurs due to a specific P01375 REA and P15056 REA interaction because P01375 REA does not prevent cell death in the presence of cisplatin , nitrogen mustard or thapsigargin . Furthermore , the cytokines P48023 REA , P50591 REA , interleukin ( IL ) - 1 , and P05231 REA do not prevent cell death when P15056 REA signaling is inhibited . The survival mechanism requires nuclear factor-kappaB ( NF-kappaB ) transcription factor activity , which is strongly induced by P01375 REA in these cells . These findings suggest that drugs that target the P15056 REA / MEK pathway could be combined with agents that target P01375 REA and / or NF-kappaB signaling to provide exciting new therapeutic opportunities for the treatment of melanoma .

Flow cytometric analysis of mammalian glial cultures treated with methotrexate . DB00563 MEN ( MTX ) is an antineoplastic drug that acts by competitive inhibition of the enzyme dihydrofolate reductase ( P00374 REA ) . MTX treatment of cultured cell lines leads to the emergence of resistant cell populations . Studies using stepwise selection procedures have demonstrated that MTX resistance conferred by overproduction of P00374 REA can be caused by P00374 REA gene amplification . We examined the effect of MTX on cells whose origin more closely approximates the in vivo condition by developing a culture system using dissociated brain tissue from 17-19 day old mouse embryos . At the first passage , cultures were divided into control and MTX groups . Cells were treated with the same or successively higher concentrations of MTX at each passage over a 3-4 month period . The first passage eliminated neurons and left a glial culture comprised of approximately 90 % astrocytes . We used the Fluorescence Activated Cell Sorter in conjunction with fluorescent dyes to measure P00374 REA content , DNA content , size , and viability of glial cells following MTX treatment . MTX-treated cells divided but grew more slowly and were larger than untreated cells . Stepwise selection in 30/60 / 90 nM or 60/120 nM MTX resulted in significant two - to threefold increases in fluorescence , and hence P00374 REA levels . Slot hybridizations assays demonstrated a threefold increase in P00374 REA gene copy number in the DNA from the 30/60 / 90 cultures . Thus , our findings were consistent with the results obtained from somatic cell lines , and lend support to the hypothesis that gene amplification may be a common mechanism for the acquisition of resistance in many types of cells . They also indicate that glial cells may be a specific target for cytotoxic effects of MTX on the central nervous system .

A guide to picking the most selective kinase inhibitor tool compounds for pharmacological validation of drug targets . To establish the druggability of a target , genetic validation needs to be supplemented with pharmacological validation . Pharmacological studies , especially in the kinase field , are hampered by the fact that many reference inhibitors are not fully selective for one target . Fortunately , the initial trickle of selective inhibitors released in the public domain has steadily swelled into a stream . However , rationally picking the most selective tool compound out of the increasing amounts of available inhibitors has become progressively difficult due to the lack of accurate quantitative descriptors of drug selectivity . A recently published approach , termed ' selectivity entropy ' , is an improved way of expressing selectivity as a single-value parameter and enables rank ordering of inhibitors . We provide a guide to select the best tool compounds for pharmacological validation experiments of candidate drug targets using selectivity entropy . In addition , we recommend which inhibitors to use for studying the biology of the 20 most investigated kinases that are clinically relevant : Abl ( P00519 REA ) , P31749 REA , Q9UM73 , Aurora A / B , CDKs , MET , P07333 REA ( P07333 REA ) , P00533 REA , P36888 REA , P04626 REA ( P04626 REA ) , O14920 REA ( O14920 REA ) , O60674 REA / 3 , P45983 REA / 2/3 ( P45983 REA / 9/10 ) , Q02750 REA / 2 , P53350 REA , PI3Ks , p38α ( Q16539 REA ) , P15056 REA , P12931 REA and P35968 REA ( P35968 REA ) .

A novel function for platelet-derived growth factor D : induction of osteoclastic differentiation for intraosseous tumor growth . Although increasing evidence suggests a critical role for platelet-derived growth factor ( PDGF ) receptor β ( β - P09619 REA ) signaling in prostate cancer ( PCa ) progression , the precise roles of β - P09619 REA and PDGF isoform-specific cell signaling have not been delineated . Recently , we identified the Q9GZP0 isoform as a ligand for β - P09619 REA in PCa and showed that Q9GZP0 is activated by serine protease-mediated proteolytic removal of the CUB domain in a two-step process , yielding first a hemidimer ( HD ) and then a growth factor domain dimer . Herein , we demonstrate that the expression of Q9GZP0 in human PCa LNCaP cells leads to enhanced bone tumor growth and bone responses in immunodeficient mice . Histopathological analyses of bone tumors generated by Q9GZP0 - expressing LNCaP cells ( LNCaP - Q9GZP0 ) revealed osteolytic and osteoblastic responses similar to those observed in human PCa bone metastases . Importantly , we discovered a novel function of Q9GZP0 in the regulation of osteoclast differentiation , independent of the O14788 REA / Q9Y6Q6 REA signaling axis . Although both PDGF-B and - D were able to activate β - P09619 REA , only Q9GZP0 was able to induce osteoclastic differentiation in vitro , and upregulate the expression and nuclear translocation of nuclear factor of activated T cells 1 , a master transcription factor for osteoclastogenesis . Taken together , these results reveal a new function of Q9GZP0 as a regulator of osteoclastic differentiation , an activity critical for the establishment of skeletal metastatic deposit in PCa patients .

Transduction of P28906 REA + cells with lentiviral vectors enables the production of large quantities of transgene-expressing immature and mature dendritic cells . BACKGROUND : Genetically engineered dendritic cells ( DC ) presenting specific antigens to T cells may be of great interest for immunotherapy . For this reason , the production of transgene-expressing DC derived from P28906 REA + cells transduced either shortly after ex vivo purification or during their differentiation into DC were evaluated . METHODS : P28906 REA + cells were transduced with lentivectors encoding for GFP before or after 21 days of culture with P36888 REA - ligand , thrombopoietin and stem cell factor and induction into DC with GM - P04141 REA + P05112 REA ( G4 ) or G4 + P01375 REA ( GT4 ) . GFP and DC-specific marker expression was assessed by flow cytometry , and allostimulatory capacity was evaluated on GFP + and GFP - sorted cells . RESULTS : Immature ( G4 - induced ) DC obtained from amplified P28906 REA + cells were transducible by lentiviral vectors while mature ( GT4 - induced ) DC were rather refractory . Moreover , since differentiated DC did not proliferate , large quantities of vectors were required to generate transgene-expressing cells with this protocol . In contrast , greater numbers of both immature and mature GFP - expressing DC were obtained with P28906 REA + cells exposed to lentivector shortly after purification . By the time of DC induction , GFP + cells had increased by approximately 170 - fold . After DC induction with G4 , 32 % of CD1a + , HLA-DR + , or P25942 REA + cells expressed GFP . CD1a + P12830 REA + GFP + Langerhans-like DC were also obtained . Incubation with P01375 REA induced mature Q01151 + GFP + DC that displayed a higher allostimulatory capacity than cells induced with G4 alone . CONCLUSION : The transduction of a small number of P28906 REA + cells with minimal doses of lentivector may allow for the production of a large number of DC expressing selected antigens useful for immunotherapy .

Discovery of potent , orally active compounds of tyrosine kinase and serine / threonine-protein kinase inhibitor with anti-tumor activity in preclinical assays . Traditional medicines have become the most productive source of leads for drugs development , particularly as anti-cancer agents . Various screening approaches are being applied . DB00398 SUB , a multikinase inhibitor , is used to treat primary kidney cancer ( advanced renal cell carcinoma ) and advanced primary liver cancer . A small library of compounds analogous to sorafenib were designed and screened for the treatment of liver cancer . Multiple members of the family in an assay panel of tyrosine kinase family and serine / threonine-protein kinase family , including VEGFR , Abl , Aurora A , p 38 , Lck , Src , P09619 REA , Flt 3 , c-RAF , c - P10721 REA , MEK ( MAPKK ) were selected to test these compounds . Analysis of the selectivity patterns for these compounds shows specificity for many kinase families . IC₅₀ were measured for the selected compounds . Multiple compounds have very similar kinase inhibition profiles of VEGFR , Flt 3 , FGFR to that of sorafenib . The IC₅₀ of c-RAF of BB1 is lower than sorafenib . The IC₅₀ of c-RAF of BB3 - 12 is higher than that of sorafenib . For Flt 3 , IC₅₀ of BB1 - 4 is less than sorafenib . The IC₅₀ value of P35968 REA of BB1 - 10 is less than sorafenib . especially against c-RAF , P09619 REA , c - P10721 REA , P35968 REA compared to sorafenib . These compounds are potent Raf 1 and Flt 4 kinase inhibitors .

Revealing multi-binding sites for taspine to P35968 REA by cell membrane chromatography zonal elution . A new high-expression vascular endothelial growth factor receptor - 2 ( P35968 REA ) cell membrane chromatography ( CMC ) method was developed to investigate the affinity of ligands for P35968 REA . An HEK 293 P35968 REA / CMC system was applied to specifically recognize ligands acting on P35968 REA . DB00398 SUB was used as a mobile phase additive to evaluate the effect of the marker ' s concentration on the retention of sorafenib and taspine , respectively . The relationship among the retention , the types of binding sites and the affinity of taspine binding to P35968 REA has also been concerned . The retention behavior indicated that sorafenib had two major binding regions on P35968 REA , and that taspine might act as a multi-target P35968 REA inhibitor with similar biological activity to sorafenib . The equilibrium dissociation constants ( K ( D ) ) obtained from the model are ( 5.25 ± 0.31 ) × 10 ⁻ ⁷ and ( 9.88 ± 0.54 ) × 10 ⁻ ⁵ mol L ⁻ ¹ for sorafenib at the high - and low-affinity sites , respectively , and the corresponding values for taspine are ( 3.88 ± 0.31 ) × 10 ⁻ ⁶ and ( 7.04 ± 0.49 ) × 10 ⁻ ⁵ mol L ⁻ ¹ . The two types of binding sites contributed about a 1:2 ratio on the retention of taspine .

Connexin 43 and P29323 REA regulate tension-induced signal transduction in human periodontal ligament fibroblasts . Periodontal ligament ( PDL ) fibroblasts play an important role in preserving periodontal homeostasis and transmitting mechanical signals to alveolar bone . Connexin 43 ( P17302 REA ) , a gap junction protein , is essential for bone homeostasis and regulates bone remodeling . However , the function of P17302 REA in human PDL fibroblast-regulated bone remodeling has not yet been elucidated . In this study , human PDL fibroblasts were exposed to cyclic mechanical tension with a maximum 5 % elongation for different durations . We then examined the expression of signaling molecules related to osteogenesis and osteoclastogenesis at both the mRNA and protein levels as well as the activity of extracellular signal-regulated kinase ( P29323 REA ) in human PDL fibroblasts after loading . We found that mechanical tension increased P17302 REA , which further upregulated osteogenic ( e . g . , Q13950 REA , Osterix , and O00300 REA ) and down-regulated osteoclastogenic ( e . g . , O14788 REA ) signaling molecules . Suppressing P17302 REA gene ( Gja 1 ) by siRNA inhibited the increase in osteogenesis-related molecules but enhanced O14788 REA expression . Similar to P17302 REA , activated P27361 REA / 2 was also enhanced by mechanical tension and suppressed by P17302 REA siRNA . Inhibition of P27361 REA / 2 signaling using PD98059 reduced the tension-regulated increase in osteogenesis-related molecules but enhanced that of osteoclastogenesis-related ones . These findings suggest that cyclic tension may involve into the osteogenic or osteoclastogenetic differentiation potential of human PDL fibroblasts via the P17302 REA - P27361 REA / 2 signaling pathway .

Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 REA ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism ( PE ) . tPA has not been replaced by third generation plasminogen activators , e . g . DB00015 ( Ret ) and DB00031 MEN ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor - 1 ( e . g . P05121 REA ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 REA than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility .