MH_dev_35

Suppression of androgen receptor-mediated gene expression by a sequence-specific DNA-binding polyamide . P10275 ( 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 .

Predictive factors for response to docetaxel in human breast cancers . DB01248 SUB has come into wide use recently for the treatment of breast cancer in neoadjuvant , adjuvant and metastatic settings . DB01248 SUB binds to beta-tubulin and causes kinetic abnormalities in the dynamics of microtubules by increasing their polymerization and inhibiting their depolymerization , resulting in elevated levels of microtubule formation . During metaphase , defective spindle formation induced by docetaxel activates the mitotic checkpoint and leads to cell cycle arrest , culminating in apoptosis . However , docetaxel is not effective for all breast cancers . For example , in metastatic settings , the response rate to docetaxel reportedly ranges from 30 to 50 % . It is therefore very important to develop a diagnostic method with high accuracy for the prediction of sensitivity to docetaxel in order to avoid unnecessary treatment . Currently it is impossible to identify , before the initiation of therapy , the patients for whom docetaxel will be effective . Various biological parameters have been studied clinically for their ability to predict response to docetaxel , such as parameters related to : ( 1 ) efflux ( p-glycoprotein ) and metabolism ( P08684 ) ; ( 2 ) beta-tubulin ( somatic mutation of beta-tubulin and changes in beta-tubulin isotypes levels ) ; ( 3 ) cell cycle ( P04626 , P38398 and Aurora-A ) ; and ( 4 ) apoptosis ( p53 , P10415 and thioredoxin ) . More recently , gene expression profiling techniques have been used for the development of a prediction model for response to docetaxel . In the present paper , clinical studies that have been conducted recently to identify predictive factors for response to docetaxel are reviewed together with a presentation of our recent work in this field .

Effect of kinase inhibitors on the therapeutic properties of monoclonal antibodies . Targeted therapies of malignancies currently consist of therapeutic monoclonal antibodies and small molecule kinase inhibitors . The combination of these novel agents raises the issue of potential antagonisms . We evaluated the potential effect of 4 kinase inhibitors , including the Q06187 inhibitor ibrutinib , and 3 PI3K inhibitors idelalisib , DB00238 - BEZ 235 and LY294002 , on the effects of the 3 monoclonal antibodies , rituximab and obinutuzumab ( directed against P11836 ) and trastuzumab ( directed against P04626 ) . We found that ibrutinib potently inhibits antibody-dependent cell-mediated cytotoxicity exerted by all antibodies , with a 50 % inhibitory concentration of 0.2 microM for trastuzumab , 0.5 microM for rituximab and 2 microM for obinutuzumab , suggesting a lesser effect in combination with obinutuzumab than with rituximab . The 4 kinase inhibitors were found to inhibit phagocytosis by fresh human neutrophils , as well as antibody-dependent cellular phagocytosis induced by the 3 antibodies . Conversely co-administration of ibrutinib with rituximab , obinutuzumab or trastuzumab did not demonstrate any inhibitory effect of ibrutinib in vivo in murine xenograft models . In conclusion , some kinase inhibitors , in particular , ibrutinib , are likely to exert inhibitory effects on innate immune cells . However , these effects do not compromise the antitumor activity of monoclonal antibodies in vivo in the models that were evaluated .

Synthesis and biological evaluation of novel pyrrolidine -2,5- dione derivatives as potential antidepressant agents . Part 1 . A series of 3 - ( 1H - indol - 3 - yl ) pyrrolidine -2,5- dione derivatives was synthesized and their biological activity was evaluated . The chemical structures of the newly prepared compounds were confirmed by ( 1 ) H NMR , ( 13 ) C NMR and P19957 - HRMS spectra data . All tested compounds proved to be potent P08908 receptor and serotonin transporter protein ( P31645 ) ligands . Among them , compounds 15 , 18 , 19 and 30 showed significant affinity for P08908 and P31645 . Computer docking simulations carried out for compounds 15 , 31 and 32 to models of P08908 receptor and P31645 confirm the results of biological tests . Due to high affinity for the P08908 receptor and moderate affinity for P31645 , compounds 31 , 32 , 35 , and 37 were evaluated for their affinity for D2L , P50406 , P34969 and 5 - Q13049 receptors . In vivo tests , in turn , resulted in determining the functional activity of compounds 15 , 18 , 19 and 30 to the P08908 receptor . The results of these tests indicate that all of the ligands possess properties characteristic of P08908 receptor agonists .

Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 - mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) - derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 - mediated signaling . The crosstalk of P35367 - mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 - α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 - α and P05231 production of BM-DCs . P35367 - specific agonists were able to enhance P01375 - α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 - α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 - mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 - mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation .

The cooperation between hMena overexpression and P04626 signalling in breast cancer . hMena and the epithelial specific isoform hMena ( 11a ) are actin cytoskeleton regulatory proteins belonging to the Ena / P50552 family . P01133 treatment of breast cancer cell lines upregulates hMena / hMena ( 11a ) expression and phosphorylates hMena ( 11a ) , suggesting cross-talk between the ErbB receptor family and hMena / hMena ( 11a ) in breast cancer . The aim of this study was to determine whether the hMena / hMena ( 11a ) overexpression cooperates with HER - 2 signalling , thereby affecting the P04626 mitogenic activity in breast cancer . In a cohort of breast cancer tissue samples a significant correlation among hMena , P04626 overexpression , the proliferation index ( high Ki67 ) , and phosphorylated MAPK and AKT was found and among the molecular subtypes the highest frequency of hMena overexpressing tumors was found in the P04626 subtype . From a clinical viewpoint , concomitant overexpression of P04626 and hMena identifies a subgroup of breast cancer patients showing the worst prognosis , indicating that hMena overexpression adds prognostic information to P04626 overexpressing tumors . To identify a functional link between P04626 and hMena , we show here that P04626 transfection in MCF 7 cells increased hMena / hMena ( 11a ) expression and hMena ( 11a ) phosphorylation . On the other hand , hMena / hMena ( 11a ) knock-down reduced P21860 , AKT and Q8TCB0 / 42 MAPK phosphorylation and inhibited the P01133 and Q02297 - dependent P04626 phosphorylation and cell proliferation . Of functional significance , hMena / hMena ( 11a ) knock-down reduced the mitogenic activity of P01133 and Q02297 . Collectively these data provide new insights into the relevance of hMena and hMena ( 11a ) as downstream effectors of the ErbB receptor family which may represent a novel prognostic indicator in breast cancer progression , helping to stratify patients .

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 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 .

DB00563 induces apoptosis through p53 / P38936 - dependent pathway and increases P12830 expression through downregulation of HDAC / Q15910 . DB00563 ( MTX ) is a dihydrofolate reductase ( P00374 ) inhibitor widely used as an anticancer drug in different kinds of human cancers . Here we investigated the anti-tumor mechanism of MTX against non-small cell lung cancer ( NSCLC ) A549 cells . MTX not only inhibited in vitro cell growth via induction of apoptosis , but also inhibited tumor formation in animal xenograft model . RNase protection assay ( RPA ) and RT-PCR demonstrated its induction of p53 target genes including DR5 , P38936 , Puma and Noxa . Moreover , MTX promoted p53 phosphorylation at Ser 15 and acetylaion at Lys 373/382 , which increase its stability and expression . The apoptosis and inhibition of cell viability induced by MTX were dependent on p53 and , partially , on P38936 . In addition , MTX also increased P12830 expression through inhibition of histone deacetylase ( HDAC ) activity and downregulation of polycomb group protein enhancer of zeste homologue 2 ( Q15910 ) . Therefore , the anticancer mechanism of MTX acts through initiation of p53 - dependent apoptosis and restoration of P12830 expression by downregulation of HDAC / Q15910 .

Discovery and structure-activity relationship of ( 1R ) - 8 - chloro -2,3 , 4,5- tetrahydro - 1 - methyl - 1H - 3 - benzazepine ( DB04871 MEN ) , a selective serotonin P28335 receptor agonist for the treatment of obesity . The synthesis and SAR of a novel 3 - benzazepine series of P28335 agonists is described . Compound 7d ( lorcaserin , APD 356 ) was identified as one of the more potent and selective compounds in vitro ( pEC 50 values in functional assays measuring [ ( 3 ) H ] phosphoinositol turnover : P28335 = 8.1 ; 5 - Q13049 = 6.8 ; P41595 = 6.1 ) and was potent in an acute in vivo rat food intake model upon oral administration ( ED50 at 6 h = 18 mg / kg ) . DB04871 MEN was further characterized in a single-dose pharmacokinetic study in rat ( t1 / 2 = 3.7 h ; F = 86 % ) and a 28 - day model of weight gain in growing Sprague-Dawley rat ( 8.5 % decrease in weight gain observed at 36 mg / kg b . i . d . ) . DB04871 MEN was selected for further evaluation in clinical trials for the treatment of obesity .

Rapid modulation of spine morphology by the 5 - Q13049 serotonin receptor through kalirin - 7 signaling . The 5 - HT ( 2A ) serotonin receptor is the most abundant serotonin receptor subtype in the cortex and is predominantly expressed in pyramidal neurons . The 5 - HT ( 2A ) receptor is a target of several hallucinogens , antipsychotics , anxiolytics , and antidepressants , and it has been associated with several psychiatric disorders , conditions that are also associated with aberrations in dendritic spine morphogenesis . However , the role of 5 - HT ( 2A ) receptors in regulating dendritic spine morphogenesis in cortical neurons is unknown . Here we show that the 5 - HT ( 2A ) receptor is present in a subset of spines , in addition to dendritic shafts . It colocalizes with P78352 and with multiple PDZ protein - 1 ( O75970 ) in a subset of dendritic spines of rat cortical pyramidal neurons . O75970 is enriched in postsynaptic density ( A5PKW4 ) fractions , is targeted to spines in pyramidal neurons , and enhances the localization of 5 - HT ( 2A ) receptors to the cell periphery . 5 - HT ( 2A ) receptor activation by the 5 - HT ( 2 ) receptor agonist DOI induced a transient increase in dendritic spine size , as well as phosphorylation of P38936 - activated kinase ( PAK ) in cultured cortical neurons . PAK is a downstream target of the neuronal Rac guanine nucleotide exchange factor ( RacGEF ) kalirin - 7 that is important for spine remodeling . O60229 - 7 regulates dendritic spine morphogenesis in neurons but its role in neuromodulator signaling has not been investigated . We show that peptide interference that prevents the localization of kalirin - 7 to the postsynaptic density disrupts DOI-induced PAK phosphorylation and spine morphogenesis . These results suggest a potential role for serotonin signaling in modulating spine morphology and kalirin - 7 ' s function at cortical synapses .

FTY 720 induces apoptosis of chronic myelogenous leukemia cells via dual activation of O43521 and P55957 and overcomes various types of resistance to tyrosine kinase inhibitors . PP2A activator FTY 720 has been shown to possess the anti-leukemic activity for chronic myelogenous leukemia ( CML ) , however , the cell killing mechanism underlying its anti-leukemic activity has remained to be verified . We investigated the precise mechanisms underlying the apoptosis induction by FTY 720 , especially focusing on the roles of BH3 - only proteins , and the therapeutic potency of FTY 720 for CML . Enforced expression of either P10415 or the dominant-negative protein of Q13158 ( Q13158 . DN ) partly protected CML cells from apoptosis by FTY 720 , indicating the involvement of both cell extrinsic and intrinsic apoptosis pathways . FTY 720 activates pro-apoptotic BH3 - only proteins : O43521 , which is essential for apoptosis by P11274 - P00519 tyrosine kinase inhibitors ( TKIs ) , and P55957 , which accelerates the extrinsic apoptosis pathway . Gene knockdown of either O43521 or P55957 partly protected K562 cells from apoptosis by FTY 720 , but the extent of cell protection was not as much as that by overexpression of either P10415 or Q13158 . DN . Moreover , knockdown of both O43521 and P55957 did not provide additional protection compared with knockdown of only O43521 or P55957 , indicating that O43521 and P55957 complement each other in apoptosis by FTY 720 , especially when either is functionally impaired . FTY 720 can overcome TKI resistance caused by P00519 kinase domain mutations , dysfunction of O43521 resulting from gene deletion polymorphism , and galectin - 3 overexpression . In addition , DB05764 , a BH3 - mimetic , significantly augmented cell death induction by FTY 720 both in TKI-sensitive and - resistant leukemic cells . These results provide the rationale that FTY 720 , with its unique effects on O43521 and P55957 , could lead to new therapeutic strategies for CML .

Initial responses of osteoblasts derived from human alveolar bone to various compressive forces . Mechanical stress generated by orthodontic force is recognized as a major factor in the modulation of alveolar bone remodeling . During this process , osteoblasts play a crucial role , not only by participating in bone formation but also by promoting osteoclastogenesis . The aim of this study was to investigate how continuous compressive force ( CF ) affects human primary osteoblasts ( HOBs ) in terms of cell proliferation , apoptosis , and expression of interleukin - 6 ( P05231 ) and chemokine CXC ligand 8 ( P10145 ) . Human primary osteoblasts , isolated from human mandibular bone pieces , were cultured with or without CF ( 1-4 g cm ( - 2 ) ) for up to 72 h . Cell viability and proliferation were evaluated using the MTT assay . RT-PCR was used to determine the levels of expression of KI67 ( a proliferation marker ) , Q07812 ( a pro-apoptotic marker ) , P10415 ( an apoptotic inhibitor ) , P05231 , and P10145 mRNAs , while a multiplexed bead immunoassay was used to measure the release of P05231 and P10145 . The results revealed that CF decreased cell viability and proliferation in a time - and force-dependent manner . After applying CF for 24 h , the mRNA expression of KI67 was markedly inhibited , whereas the mRNA expression of Q07812 and P10415 was unaltered . In addition , CF enhanced the levels of P05231 and P10145 mRNAs in a force-dependent manner , whereas the levels of the corresponding proteins were reduced in the compressed HOBs .

Apoptotic markers in a prostate cancer cell line : effect of ellagic acid . Ellagic acid ( EA ) inhibits cell growth and induces apoptosis in cultured cells ; however , the precise molecular mechanism involved in EA-induced apoptosis in prostate cancer cells is unknown . The aim of the present study was to delineate possible apoptotic pathway ( s ) involved in the EA-mediated chemotherapeutic effects in the LNCaP human prostatic cancer cell line . EA produced anti-proliferative effects through inhibition of rapamycin ( P42345 ) activation and a reduction in intracellular levels of β-catenin . Moreover , we demonstrated that EA induced apoptosis via downregulation of the anti-apoptotic proteins , silent information regulator 1 ( Q96EB6 ) , human antigen R ( Q15717 ) and heme oxygenase - 1 ( P09601 ) . EA modulated the expression of apoptosis-inducing factor ( O95831 ) resulting in a significant increase in reactive oxygen species ( ROS ) levels and the activation of caspase - 3 . Finally , we demonstrated that EA reduced both transforming growth factor-β ( TGF-β ) and interleukin - 6 ( P05231 ) levels . EA treatment resulted in the increased expression of the tumor suppressor protein P38936 and increased the percentage of apoptotic cells . In conclusion , the results suggest that EA treatment represents a new and highly effective strategy in reducing prostate cancer carcinogenesis .

Combination of ibrutinib with ABT - 199 : synergistic effects on proliferation inhibition and apoptosis in mantle cell lymphoma cells through perturbation of Q06187 , AKT and P10415 pathways .

Molecular targeting of the oncoprotein P53350 in pediatric acute myeloid leukemia : RO3280 , a novel P53350 inhibitor , induces apoptosis in leukemia cells . P53350 ( P53350 ) is highly expressed in many cancers and therefore a biomarker of transformation and potential target for the development of cancer-specific small molecule drugs . RO3280 was recently identified as a novel P53350 inhibitor ; however its therapeutic effects in leukemia treatment are still unknown . We found that the P53350 protein was highly expressed in leukemia cell lines as well as 73.3 % ( 11/15 ) of pediatric acute myeloid leukemia ( AML ) samples . P53350 mRNA expression was significantly higher in AML samples compared with control samples ( 82.95 ± 110.28 vs . 6.36 ± 6.35 ; p < 0.001 ) . Kaplan-Meier survival analysis revealed that shorter survival time correlated with high tumor P53350 expression ( p = 0.002 ) . The 50 % inhibitory concentration ( IC50 ) of RO3280 for acute leukemia cells was between 74 and 797 nM . The IC50 of RO3280 in primary acute lymphocytic leukemia ( ALL ) and AML cells was between 35.49 and 110.76 nM and 52.80 and 147.50 nM , respectively . RO3280 induced apoptosis and cell cycle disorder in leukemia cells . RO3280 treatment regulated several apoptosis-associated genes . The regulation of P43146 , P38936 , Q06187 , and O14508 was verified by western blot . These results provide insights into the potential use of RO3280 for AML therapy ; however , the underlying mechanisms remain to be determined .

P13688 impedes thyroid cancer growth but promotes invasiveness : a putative mechanism for early metastases . P13688 , also known as biliary glycoprotein ( BGP ) , CD66a , Q00839 and C - P62158 , is a member of the P06731 immunoglobulin superfamily . P13688 is a putative tumor suppressor based on diminished expression in some solid neoplasms such as colorectal carcinoma . However , P13688 is overexpressed in some tumors such as non-small cell lung cancer . To clarify the mechanism of action of this cell adhesion molecule , we studied thyroid carcinoma that has a spectrum of morphologies and variable behavior allowing separation of proliferation from invasion and metastasis . P13688 is expressed in thyroid carcinoma cell lines derived from tumors that exhibit aggressive behavior . Introduction of P13688 into endogenously deficient WRO cells resulted in reduced cell cycle progression associated with P38936 upregulation and diminished Rb phosphorylation . Forced P13688 expression enhanced cell-matrix adhesion and migration and promoted tumor invasiveness . Conversely , small interfering RNA ( siRNA ) - mediated downregulation of P13688 expression in Q9BYG7 cells accelerated cell cycle progression and significantly enhanced tumor size in xenografted mice . P13688 is not appreciably expressed in normal thyroid tissue or benign thyroid tumors . In a human thyroid tissue array , P13688 reactivity was associated with metastatic spread but not with increased tumor size . These findings identify P13688 as a unique mediator that restricts tumor growth whereas increasing metastatic potential . Our data highlight a complex repertoire of actions providing a putative mechanism underlying the spectrum of biologic behaviors associated with thyroid cancer .

Inhibitors of Q06187 and Q08881 : state of the new drugs for cancer , autoimmunity and inflammatory diseases . Q06187 and Q08881 are cytoplasmic tyrosine kinases of crucial importance for B and T cell development , with loss-of-function mutations causing X-linked agammaglobulinemia and susceptibility to severe , frequently lethal , Epstein-Barr virus infection , respectively . Over the last few years , considerable efforts have been made in order to develop small-molecule inhibitors for these kinases to treat lymphocyte malignancies , autoimmunity or allergy / hypersensitivity . The rationale is that even if complete lack of Q06187 or Q08881 during development causes severe immunodeficiency , inactivation after birth may result in a less severe phenotype . Moreover , therapy can be transient or only partially block the activity of Q06187 or Q08881 . Furthermore , a drug-induced B cell deficiency is treatable by gamma globulin substitution therapy . The newly developed Q06187 inhibitor P05154 - 32765 , recently renamed DB09053 MEN , has already entered several clinical trials for various forms of non-Hodgkin lymphoma as well as for multiple myeloma . Experimental animal studies have demonstrated highly promising treatment effects also in autoimmunity . Q08881 inhibitors are still under the early developmental phase , but it can be expected that such drugs will also become very useful . In this study , we present Q06187 and Q08881 with their signalling pathways and review the development of the corresponding inhibitors .

Simultaneous inhibition of MEK and P11802 leads to potent apoptosis in human melanoma cells . ABSTRACT Deregulation of DB01367 - RAF-MEK - P29323 and P42771 - cycylin D: P11802 / 6 - RB pathways is important for melanoma development . Chemotherapeutic agents targeting both pathways were developed but results of clinical studies with monotherapies were disappointing . We examined the effect of co-targeting both pathways with MEK inhibitor PD98059 and P11802 inhibitor 219476 on human melanoma cells lines , and found that combinatorial treatment dramatically increased apoptosis compared to the single agent treatment . The apoptosis was associated with downregulation of P10415 , Q07817 , O15392 , and upregulation of O43521 . Our results indicate that simultaneously targeting P29323 and RB pathways is a promising strategy for melanoma treatment and should encourage further in-depth investigations .

Genetic expression profiles and chromosomal alterations in sporadic breast cancer in Mexican women . Breast cancer is the second-leading cause of death among Mexican women > 35 years of age . At the molecular level , changes in many genetic pathways have been reported to be associated with this neoplasm . To analyze these changes , we determined gene expression profiles and chromosomal structural alterations in tumors from Mexican women . We obtained mRNA to identify expression profiles with microarray technology , and DNA to determine amplifications and deletions , in 10 fresh sporadic breast tumor biopsies without treatment , as well as in 10 nonaffected breast tissues . Expression profiles were compared with genetic changes observed by comparative genomic hybridization ( CGH ) . We compared the expression profiles against the structural alterations from the studied genes by means of microarrays ; at least 17 of these genes correlated with DNA copy number alterations . We found that the following genes were overexpressed : P11047 , Q07002 , P24863 , P24385 , P11487 , Q00537 , L1CAM , P21810 , and Q9ULL4 ( alias PLEXR ) . Underexpressed genes included P55211 , P09769 , O15350 , P98160 , and P07992 ; genes turned off included P42345 , P29317 ( previously P29317 ) , P29459 , Q15329 , O14763 , O00220 , Q15768 , and P10415 . The results will allow us , in the near future , to outline genes that could serve as diagnostic , prognostic , or target therapy markers for the Mexican population .

P41134 enhances docetaxel cytotoxicity in prostate cancer cells through inhibition of P38936 . To identify potential mechanisms underlying prostate cancer chemotherapy response and resistance , we compared the gene expression profiles in high-risk human prostate cancer specimens before and after neoadjuvant chemotherapy and radical prostatectomy . Among the molecular signatures associated with chemotherapy , transcripts encoding inhibitor of DNA binding 1 ( P41134 ) were significantly upregulated . The patient biochemical relapse status was monitored in a long-term follow-up . Patients with P41134 upregulation were found to be associated with longer relapse-free survival than patients without P41134 increase . This in vivo clinical association was mechanistically investigated . The chemotherapy-induced P41134 upregulation was recapitulated in the prostate cancer cell line LNCaP . DB01248 SUB dose-dependently induced P41134 transcription , which was mediated by P41134 promoter E-box chromatin modification and c-Myc binding . Stable P41134 overexpression in LNCaP increased cell proliferation , promoted G ( 1 ) cell cycle progression , and enhanced docetaxel-induced cytotoxicity . These changes were accompanied by a decrease in cellular mitochondria content , an increase in P10415 phosphorylation at serine 70 , caspase - 3 activation , and poly ( ADP-ribose ) polymerase cleavage . In contrast , P41134 siRNA in the LNCaP and C42B cell lines reduced cell proliferation and decreased docetaxel-induced cytotoxicity by inhibiting cell death . P41134 - mediated chemosensitivity enhancement was in part due to P41134 suppression of P38936 . Overexpression of P38936 in LNCaP - P41134 - overexpressing cells restored the P38936 level and reversed P41134 - enhanced chemosensitivity . These molecular data provide a mechanistic rationale for the observed in vivo clinical association between P41134 upregulation and relapse-free survival . Taken together , it shows that P41134 expression has a novel therapeutic role in prostate cancer chemotherapy and prognosis .

5 - Q9H205 - and P28335 - antagonist properties of cyamemazine : significance for its clinical anxiolytic activity . RATIONALE : DB09000 is a neuroleptic compound which possesses anxiolytic properties in humans . On the other hand , 5 - Q9H205 - and P28335 - receptors have been implicated in anxiety disorders and a previous binding study has shown that cyamemazine possesses high affinity for both serotonin receptor types . OBJECTIVE : The present study was undertaken to establish whether cyamemazine antagonizes 5 - Q9H205 - and / or P28335 - mediated responses , and whether it compares with reference compounds . METHODS : DB09000 was tested for its ability to antagonize : ( i ) 5 - Q9H205 - dependent contraction of the isolated guinea-pig ileum and bradycardic responses in the rat and ( ii ) P28335 - dependent phospholipase C ( P98160 ) stimulation in rat brain membranes . RESULTS : In isolated guinea-pig ileum , cyamemazine potently and competitively antagonized 5 - HT-dependent contractions ( pA2 = 7.52 + / - 0.08 ; n = 5 ) . In this test , cyamemazine was 5-7 times more potent ( pIC 50 = 6.75 + / - 0.13 ) than tropisetron ( pIC 50 = 6.02 + / - 0.04 ) . In rats , cyamemazine i . v . antagonized 5 - HT-dependent bradycardic responses with ID50 % = 3.2 + / - 1.5 mg / kg ( n = 4 ) . Finally , in rat brain membranes cyamemazine antagonized P28335 - dependent P98160 stimulation with Ki = 424 nM ( mianserin exhibits a Ki = 113 nM ) . CONCLUSIONS : DB09000 behaves as an antagonist at both 5 - Q9H205 - and P28335 - receptors , which compares well with reference compounds . These 5 - Q9H205 - and P28335 - antagonistic actions of cyamemazine can be involved , at least in part , in its beneficial therapeutic actions in anxiety disorders .

EPO receptor circuits for primary erythroblast survival . EPO functions primarily as an erythroblast survival factor , and its antiapoptotic actions have been proposed to involve predominantly P19957 - kinase and BCL-X pathways . Presently , the nature of EPO-regulated survival genes has been investigated through transcriptome analyses of highly responsive , primary bone marrow erythroblasts . Two proapoptotic factors , Bim and FoxO 3a , were rapidly repressed not only via the wild-type P19235 , but also by PY-deficient knocked-in P19235 alleles . In parallel , Pim 1 and Pim 3 kinases and Irs 2 were induced . For this survival gene set , induction failed via a PY-null P19235 - HM allele , but was restored upon reconstitution of a PY343 P42229 - binding site within a related P19235 - H allele . Notably , P19235 - HM supports erythropoiesis at steady state but not during anemia , while P19235 - H exhibits near wild-type P19235 activities . P19235 - H and the wild-type P19235 ( but not P19235 - HM ) also markedly stimulated the expression of Trb 3 pseudokinase , and intracellular serpin , Serpina - 3G . For SERPINA - 3G and TRB 3 , ectopic expression in EPO-dependent progenitors furthermore significantly inhibited apoptosis due to cytokine withdrawal . BCL-XL and P10415 also were studied , but in highly responsive Kit ( pos ) CD71 ( high ) Ter 119 ( neg ) erythroblasts , neither was EPO modulated . P19235 survival circuits therefore include the repression of Bim plus FoxO 3a , and P19235 / PY343 / P42229 - dependent stimulation of Pim 1 , Pim 3 , Irs 2 plus Serpina - 3G , and Trb 3 as new antiapoptotic effectors .

DB04946 MEN binding to human and rat dopamine and 5 - HT receptors . DB04946 MEN ( DB04946 MEN ; 1 - [ 4 - [ 3 - [ 4 - ( 6 - fluoro -1,2- benzisoxazol - 3 - yl ) - 1 - piperidinyl ] propoxy ] - 3 - methoxyphenyl ] ethanone ) is a compound currently in clinical trials for the treatment of schizophrenia . DB04946 MEN displays affinity for dopamine D2 receptors and for 5 - Q13049 receptors and has a variety of in vivo activities suggestive of an atypical antipsychotic . Here we present an examination of the affinity of iloperidone to a variety of human and rat homologs of dopamine and 5 - HT receptor subtypes . We employed receptor binding assays using membranes from cells stably expressing human dopamine D1 , D2S , D2L , D3 , D4 and D5 and 5 - Q13049 and P28335 receptors and rat P50406 and P34969 receptors . DB04946 MEN displayed higher affinity for the dopamine D3 receptor ( Ki = 7.1 nM ) than for the dopamine D4 receptor ( Ki = 25 nM ) . DB04946 MEN displayed high affinity for the P50406 and P34969 receptors ( Ki = 42.7 and 21.6 nM , respectively ) , and was found to have higher affinity for the 5 - Q13049 ( Ki = 5.6 nM ) than for the P28335 receptor ( Ki = 42.8 nM ) . The potential implications of this receptor binding profile are discussed in comparison with data for other antipsychotic compounds .

[ Measurement of rifampicin and clarithromycin in serum by high-performance liquid chromatography with electrochemical detection ] . DB01045 ( RFP ) induces hepatic drug-metabolizing enzymes , making drug interactions a very important clinical problem . DB01211 MEN ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 ) by RFP , so that the blood lend of P62158 decreases when RFP is administered concurrently . We connected an electrochemical detector to a high-performance liquid chromatograph ( HPLC ) for simple , rapid , easy measurement of blood concentrations of RFP and P62158 . Using samples of patient serum , normal serum , and reference standards , we compared HPLC by an external laboratory and the results of LC / MS / MS analysis with those of this new assay . A strong correlation was seen between our HPLC results and those of the external laboratory in RFP levels ( r = 0.975 , p < 0.01 ) . A strong correlation was also seen between results we obtained for P62158 with the electrochemical detector in this assay and values measured by LC / MS / MS analysis ( r = 0.995 , p < 0.01 ) . Our method enabled simple , rapid measurement of RFP and P62158 by connecting the HPLC and electrochemical detector in tandem . This system was used to modulate dosage during combined therapy with RFP and P62158 . The therapeutic effect for nontuberculous mycobacteriosis is expected to improve , and our HPLC is expected to be useful for simple , rapid , easy measurement of blood concentrations .

Gene profiling in Pap-cell smears of high-risk human papillomavirus-positive squamous cervical carcinoma . OBJECTIVE : The purpose of the study was to investigate benign and malignant squamous cervical cells obtained by cervical swabs with regard to differentially expressed genes and gene expression profiling , in order to evaluate the biological behavior and clinical outcome of cervical malignancies . METHODS : Cervical squamous cells from six women with high-risk human papillomavirus positive [ HR-HPV ( + ) ] cervical carcinoma and from six HPV-negative women with normal ectocervical cells were analyzed by cDNA array . RESULTS : cDNA over-expression of several genes such as MET ( c-met ) , Nm23 - H1 ( P15531 ) , P00533 , P21802 , Nm23 - H2 ( P22392 ) , P04626 ( c-erbB - 2 ) , cyclin-dependent kinase inhibitor 4 ( CDKN 2A , P42771 ) , cytokeratin 8 ( P05787 ) , P01116 ( K-ras ) , P17948 , KGF ( P21781 ) , P10415 - like 2 protein ( Q92843 ) , Q15303 , P04198 ( N-myc ) , cyclin D1 ( P24385 ) , P10721 ( c-kit ) , secreted phosphoprotein 1 ( P10451 ) and P42224 , was significant in cervical squamous cell carcinoma ( CSCC ) . Gene expression was downregulated for 13 genes in CSCC , such as interleukin 1 alpha ( P01583 ) , the transforming growth factor receptor beta superfamily ( TGFbeta ; P01137 ) , some members of the insulin-like growth factor binding proteins ( IGFBPs ) and the integrin family ( P23229 , P05556 ) . CONCLUSION : This study was focused on the gene expression profiling of HR-HPV ( - ) and ( + ) cervical squamous cells and CSCC obtained by cytobrush . We observed gene expression patterns and signaling pathways that permit the investigator to distinguish between benign squamous cervical cells and CSCC with and without HPV infection .

[ Innate resistance to thymidylate synthase inhibition after 5 - fluorouracil treatment - - a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MEN MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7 - days administration of DB09327 ( U - 7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U - 1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U - 7 group compared with U - 1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg / kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect .

Imatinib restores P50552 activity and its interaction with Q15942 in P11274 - P00519 leukemic cells . P50552 ( P50552 ) and Q15942 are interacting proteins involved in cellular adhesion and motility . PKA phosphorylates P50552 at serine 157 , regulating P50552 cellular functions . P50552 interacts with P00519 and is a substrate of the P11274 - P00519 oncoprotein . The presence of P11274 - P00519 protein drives oncogenesis in patients with chronic myeloid leukemia ( CML ) due to a constitutive activation of tyrosine kinase activity . However , the function of P50552 and Q15942 in P11274 - P00519 pathway and the role of P50552 in CML cells remain unknown . In vitro experiments using K562 cells showed the involvement of P50552 in P11274 - P00519 signaling . P50552 and Q15942 inhibition decreased the expression of anti-apoptotic proteins , P10415 and BCL-XL . Imatinib induced an increase in phosphorylation at Ser 157 of P50552 and decreased P50552 and P11274 - P00519 interaction . P50552 did not interact with Q15942 in K562 cells ; however , after Imatinib treatment , this interaction was restored . Corroborating our data , we demonstrated the absence of phosphorylation at Ser 157 in P50552 in the bone marrow of CML patients , in contrast to healthy donors . Phosphorylation of P50552 on Ser 157 was restored in Imatinib responsive patients though not in the resistant patients . Therefore , we herein identified a possible role of P50552 in CML pathogenesis , through the regulation of P11274 - P00519 effector proteins or the absence of phosphorylation at Ser 157 in P50552 .

Association of genetic polymorphisms with personality profile in individuals without psychiatric disorders . OBJECTIVE : Population-based twin studies demonstrate that approximately 40-50 % of the variability in personality dimensions results from genetic factors . This study assessed selected polymorphisms in the P21964 Val 158Met , P21397 3 ' VNTR , 5HTTLPR , 102T / C 5 - Q13049 , Q01959 3 ' VNTR and P14416 exon 8 genes and evaluated their association with personality profiles , anxiety levels , and depressiveness in healthy subjects . METHODS : This study included 406 unrelated ( mean age 38.51 years ) , mentally and somatically healthy Caucasian subjects of Polish origin . The prevalence of the gene variants mentioned above and their association with personality profiles , anxiety levels , and depressiveness was assessed using the Temperament and Character Inventory , NEO Five-Factor Inventory , Spielberger ' s State-Trait Anxiety Inventory and Beck ' s Depression Inventory . RESULTS : The effects of the 5HTTLPR gene on the s / s genotype and empathy ( P06681 ) were lowest in the entire group . The effects of gender , age and the Q13049 gene for the T / T genotype and attachment ( Q7Z3Z2 ) were highest in women . The effects of gender , age and the Q01959 gene on the 9/9 Q01959 genotype , compassion ( C4 ) and cooperativeness ( C ) were lowest in women . The effects of gender , age and the P21964 gene on the DB00134 / DB00134 genotype and neuroticism ( P04626 ) NEO-FFI were also lowest in women . CONCLUSIONS : Our results suggest considerable influence of individual genes on the formation of personality traits .

Hypoxic / normoxic preconditioning increases endothelial differentiation potential of human bone marrow CD133 + cells . CD133 + cells are hemangioblasts that have capacity to generate into both hematopoietic and endothelial cells ( ECs ) . Hypoxia / normoxia has shown to be the regulator of the balance between stemness and differentiation . In this study we performed Agilent ' s whole human genome oligo microarray analysis and examined the differentiation potential of the bone-marrow-derived CD133 + cells after hypoxic / normoxic preconditioning of CD133 + cells . Results showed that there was no significant increase in erythroid colony forming unit ( CFU-E ) and CFU-granulocyte , erythrocyte , monocyte , and megakaryocyte formation with cells treated under hypoxia / normoxia . However , a significant increment of EC forming unit at 24 h ( 143.2 + / - 8.0 % ) compared to 0 h ( 100 + / - 11.4 % ) was observed in CFU-EC analysis . Reverse transcription-polymerase chain reaction and immunostaining analysis showed that the differentiated cells diminished hematopoietic stem cell surface markers and acquired the gene markers and functional phenotype of ECs . The transcriptome profile revealed a cluster of 232 downregulated and 498 upregulated genes in cells treated for 24 h under hypoxia . The upregulated genes include angiogenic genes , angiogenic growth factor genes , angiogenic cytokine and chemokine genes , as well as angiogenic-positive regulatory genes , including Q14512 , PDGFB , Q16663 , P48061 , P8 0162 , P05231 , P21246 , O14944 , P04626 , O95136 , P11487 , Q92913 , Q99988 , P05412 , L1CAM , Q02297 , P08138 , and PDGFB . On the other hand , angiogenesis inhibitors and related genes , including P29459 , P98177 , Q9NY15 , and P16035 , are downregulated . Taken together , hypoxic / normoxic preconditioning may lead to the differentiation of CD133 + cells toward endothelial lineage , which may improve the current clinical trial studies .

Placental autophagy regulation by the Q9UMX3 - Q07820 rheostat . Autophagy is the catabolic degradation of cellular cytoplasmic constituents via the lysosomal pathway that physiologically elicits a primarily cytoprotective function , but can rapidly be upregulated in response to stressors thereby inducing cell death . We have reported that the balance between the P10415 family proteins Q9UMX3 and Q07820 regulates human trophoblast cell fate and its alteration toward cell death typifies preeclampsia . Here we demonstrate that Q9UMX3 is a potent inducer of autophagy as shown by increased LC3B - II production , autophagosomal formation and lysosomal activation in P29320 293 . In contrast , using JEG 3 cells we showed that prosurvival Q07820 acts as a repressor of autophagy via an interaction with Q14457 , which is abrogated by Q9UMX3 . We found that Q07820 - cleaved products , specifically MCL 1c157 , trigger autophagy while the splicing variant MCL 1S has no effect . Treatment of JEG 3 cells with nitric oxide donor SNP resulted in Q9UMX3 - Q07820 rheostat dysregulation , favoring Q9UMX3 accumulation , thereby inducing autophagy . Overexpression of Q07820 rescued oxidative stress-induced autophagy . Of clinical relevance , we report aberrant autophagy levels in the preeclamptic placenta due to impaired recruitment of Q14457 to Q07820 . Our data provided the first evidence for a key role of the Q9UMX3 - Q07820 system in regulating autophagy in the human placenta , whereby an adverse environment as seen in preeclampsia tilts the Q9UMX3 - Q07820 balance toward the build-up of isoforms that triggers placental autophagy .

The potential role of PD0332991 ( DB09073 MEN ) in the treatment of multiple myeloma . INTRODUCTION : Multiple myeloma ( MM ) remains an incurable malignancy indicating a need for continued investigation of novel therapies . Recent studies have highlighted the role of cyclin-dependent kinases ( CDK ) in the pathogenesis of MM . PD0332991 ( DB09073 MEN ) is an orally bioavailable , highly selective inhibitor of the P11802 / 6 - cyclin complex and downstream retinoblastoma protein ( Rb ) activation pathway that induces cell cycle arrest in the P55008 phase . AREAS COVERED : In this review , the authors summarize the role of the P11802 / 6 signaling pathway in MM . They also summarize the development of PD0332991 as a specific inhibitor of P11802 / 6 , and the reported preclinical and clinical data supporting the potential role of PD0332991 in MM . EXPERT OPINION : While PD0332991 is essentially cytostatic , inducing prolonged P55008 arrest , it enhances the cytotoxic effect of other agents effective in MM , including bortezomib and lenalidomide , as confirmed in early phase clinical trials . However , with a plethora of other drugs of different classes being tested in MM , further development of PD0332991 will depend on defining the most efficacious combination with least toxicity . An unexplored opportunity remains the potential protective effect of PD0332991 against lytic bone lesions of MM . The next few years are likely to better define the place of PD0332991 in the treatment of MM .

Characterization of the inhibitory effects of erythromycin and clarithromycin on the Q12809 potassium channel . Both erythromycin and clarithromycin have been reported to cause QT prolongation and the cardiac arrhythmia torsade de pointes in humans , however direct evidence documenting that these drugs produce this effect by blocking human cardiac ion channels is lacking . The goal of this study was to test the hypothesis that these macrolide antibiotics significantly block the delayed rectifier current ( IKr ) encoded by Q12809 ( the human ether-a-go-go-related gene ) at drug concentrations , temperature and ionic conditions mimicking those occurring in human subjects . DB01345 currents in P29320 293 cells stably transfected with Q12809 were recorded using a whole cell voltage clamp method . Exposure of cells to erythromycin reduced the Q12809 encoded potassium current in a concentration dependent manner with an IC50 of 38.9 + / - 1.2 microM and Hill Slope factor of 0.4 + / - 0.1 . DB01211 MEN produced a similar concentration-dependent block with an IC50 of 45.7 + / - 1.1 microM and Hill Slope factor of 1.0 + / - 0.1 . Erythromycin ( 25-250 microM ) and clarithromycin ( 5 or 25 microM ) also produced a significant decrease in the integral of the current evoked by an action potential shaped voltage clamp protocol . The results of this study document that both erythromycin and clarithromycin significantly inhibit the Q12809 potassium current at clinically relevant concentrations .

Drug insight : Use of docetaxel in prostate and urothelial cancers . Taxanes have emerged as a potent class of chemotherapeutic agents in many malignancies , with two taxanes now in clinical use . Their mechanism of action against tumor cells is by alteration of microtubule dynamics , which causes cell-cycle arrest during mitosis . DB01248 SUB binds to the microtubules with a higher affinity than paclitaxel , and over a broader range of cell-cycle activities . It has also been shown to promote apoptosis via P10415 phosphorylation . In hormone-refractory prostate cancer , docetaxel has been studied as both a single agent and in combination with estramustine , and in different treatment schedules , with demonstrated efficacy . Two phase III trials have confirmed a survival benefit , making docetaxel the first chemotherapy agent with proven efficacy against prostate cancer . In urothelial cancer , docetaxel has demonstrated activity and has been investigated as a single agent and in combination regimens . A phase III trial comparing docetaxel and cisplatin to methotrexate , vinblastine , doxorubicin , and cisplatin was inferior when evaluating response rates and overall survival . More recent phase II trials combining docetaxel with two additional agents have shown promise , but confirmatory trials are needed .

P10275 expression in breast cancer patients tested for P38398 and P51587 mutations . AIM : To assess the expression of receptors for androgen ( AR ) , oestrogen ( ER ) and progesterone ( PR ) as well as human epidermal growth factor receptor type 2 ( Her - 2 / neu ) status of breast carcinomas in breast cancer susceptibility gene ( BRCA ) P38398 / 2 mutation carriers and P38398 / 2 negative tested women . METHODS : One hundred and thirty-five breast cancers in women tested for P38398 / 2 mutations . Screening for P38398 and P51587 mutations was performed by direct sequencing of all P38398 and P51587 exons as well as the surrounding intronic sequences . Additionally , BRCA genes were analysed with multiplex ligation-dependent probe amplification . Consecutive paraffin sections were examined immunohistochemically for AR , ER , PR and Her - 2 / neu . RESULTS : Of the 135 tumours , 43 ( 32 % ) were P38398 - related , 18 ( 13 % ) were P51587 - related and 74 ( 55 % ) were P38398 / 2 - negative . Seventy-two per cent of the P38398 - related , 22 % of the P51587 - related and 12 % of the P38398 / 2 - negative tumours were triple ( ER , PR , Her 2neu ) - negative . Eighty-four per cent of P38398 mutated cancers were high-grade ( P46379 ) tumours . ARs were expressed in 30 % ( 13 of 43 ) of P38398 - related , in 78 % ( 14 of 18 ) in P51587 - related tumours and in 76 % ( 56 of 74 ) in P38398 / 2 negative tumours . Twenty-one per cent of ER-negative P38398 - related tumours expressed androgen receptors . CONCLUSION : Approximately one in five P38398 mutated breast cancers negative for ER and PR express androgen receptors . Modulation of AR might open a new avenue for treating these high-risk cancers .

Effects of dutasteride on the expression of genes related to androgen metabolism and related pathway in human prostate cancer cell lines . Androgens play an important role in controlling the growth of the normal prostate gland and in the pathogenesis of benign prostate hyperplasia , and prostate cancer . Although testosterone is the main androgen secreted from the testes , dihydrotestosterone ( DB02901 ) , a more potent androgen converted from testosterone by 5alpha - reductase isozymes , type I and II , is the major androgen in the prostate cells . The aim of this study is to investigate the cellular and molecular effects of dutasteride , a potent inhibitor of 5alpha - reductase type I and type II , in androgen-responsive ( LNCaP ) and androgen-unresponsive ( DU145 ) human prostate cancer ( PCa ) cell lines . The expression pattern of 190 genes , selected on the basis of their proved or potential role in prostate cancerogenesis related to androgen signalling , were analysed using a low density home-made oligoarray ( AndroChip 2 ) . Our results show that dutasteride reduces cell viability and cell proliferation in both cell lines tested . AndroChip 2 gene signature identified in LNCaP a total of 11 genes differentially expressed ( FC > or = + / -1.5 ) . Eight of these genes , were overexpressed and three were underexpressed . Overexpressed genes included genes encoding for proteins involved in biosynthesis and metabolism of androgen ( P14061 ; P37058 ; P19099 ) , androgen receptor and androgen receptor co-regulators ( AR ; P24385 ) , and signal transduction ( P04626 ; V - P62158 ; Q07889 ) whereas , underexpressed genes ( KLK 3 ; P20151 ; Q15392 ) were androgen-regulated genes ( ARGs ) . No differentially expressed genes were scored in DU145 . Microarray data were confirmed by quantitative real-time PCR assay ( QRT-PCR ) . These data offer a selective genomic signature for dutasteride treatment in prostate epithelial cells and provide important insights in prostate cancer pathophysiology .

DB09073 MEN ( PD 0332991 ) : targeting the cell cycle machinery in breast cancer . INTRODUCTION : The cyclin D-cyclin-dependent kinases 4 and 6 ( P11802 / 6 ) - retinoblastoma ( P06400 ) pathway , governing the cell cycle restriction point , is frequently altered in breast cancer and is a potentially relevant target for anticancer therapy . DB09073 MENMAX DB09073 MEN ( PD 0332991 ) , a potent and selective inhibitor of P11802 and Q00534 , inhibits proliferation of several P06400 - positive cancer cell lines and xenograft models . AREAS COVERED : The basic features and abnormalities of the cell cycle in breast cancer are described , along with their involvement in estrogen signaling and endocrine resistance . The pharmacological features of palbociclib , its activity in preclinical models of breast cancer and the potential determinants of response are then illustrated , and its clinical development in breast cancer described . A literature search on the topic was conducted through PubMed and the proceedings of the main cancer congresses of recent years . EXPERT OPINION : The combination of palbociclib with endocrine agents is a very promising treatment and Phase III clinical trials are ongoing to confirm its efficacy . Further , potentially useful combinations are those with drugs targeting mitogenic signaling pathways , such as P04626 - and PI3K - inhibitors . Combination with chemotherapy seems more problematic , as antagonism has been reported in preclinical models . The identification of predictive factors , already explored in preclinical studies , must be further refined and validated in clinical trials .

' VASPFix ' for measurement of P50552 phosphorylation in platelets and for monitoring effects of Q9H244 antagonists . P50552 ( P50552 ) is phosphorylated and dephosphorylated consequent to increases and decreases in cyclic nucleotide levels . Monitoring changes in P50552 phosphorylation is an established method for indirect measurement of cyclic nucleotides . Here we describe the use of an innovative cocktail , VASPFix , which allows sensitive and reproducible measurement of phosphorylated P50552 ( P50552 - P ) in a simple , single-step procedure using cytometric bead technology . Frozen VASPFix-treated samples are stable for at least six months prior to analysis . We successfully used VASPFix to measure P50552 - P in platelets in both platelet-rich plasma and blood in response to compounds that increase ( dibutyryl DB02527 , adenosine , iloprost , PGE 1 ) and decrease ( ADP , PGE 1 ) DB02527 , and to determine the effects of certain receptor antagonists on the results obtained . The change in P50552 - P brought about by adding ADP to PGE 1 - stimulated platelets is a combination of the effect of ADP at the Q9H244 receptor and of PGE 1 at both IP and EP3 receptors . For iloprost-stimulated platelets EP3 receptors are not involved . A procedure in which iloprost , ADP and VASPFix were used to determine effectiveness of clopidogrel and prasugrel in patients was compared with an established commercial procedure that uses PGE 1 and ADP ; the latter produced higher platelet reactivity values that were the result of PGE 1 interacting with platelet EP3 receptors . We conclude that VASPFix can be used both as a research tool and for clinical investigations and provides better specificity for Q9H244 receptor inhibition . The latter confers a distinct advantage over existing methods used to monitor effects of Q9H244 antagonists on platelet function .

The P28335 receptor agonist lorcaserin reduces nicotine self-administration , discrimination , and reinstatement : relationship to feeding behavior and impulse control . DB04871 MEN ( ( 1R ) - 8 - chloro - 1 - methyl -2,3 , 4,5- tetrahydro - 1H - 3 - benzazepine HCl ) is a selective 5 - HT ( 2C ) receptor agonist with clinical efficacy in phase-III obesity trials . Based on evidence that this drug class also affects behaviors motivated by drug reinforcement , we compared the effect of lorcaserin on behavior maintained by food and nicotine reinforcement , as well as the stimulant and discriminative stimulus properties of nicotine in the rat . Acutely administered lorcaserin ( 0.3- 3 mg / kg , subcutaneous ( SC ) ) dose dependently reduced feeding induced by 22 - h food deprivation or palatability . Effects up to 1 mg / kg were consistent with a specific effect on feeding motivation . DB04871 MEN ( 0.6- 1 mg / kg , SC ) reduced operant responding for food on progressive and fixed ratio schedules of reinforcement . In this dose range lorcaserin also reversed the motor stimulant effect of nicotine , reduced intravenous self-administration of nicotine , and attenuated the nicotine cue in rats trained to discriminate nicotine from saline . DB04871 MEN also reduced the reinstatement of nicotine-seeking behavior elicited by a compound cue comprising a nicotine prime and conditioned stimulus previously paired with nicotine reinforcement . DB04871 MEN did not reinstate nicotine-seeking behavior or substitute for a nicotine cue . Finally , lorcaserin ( 0.3- 1 mg / kg ) reduced nicotine-induced increases in anticipatory responding , a measure of impulsive action , in rats performing the five-choice serial reaction time task . Importantly , these results indicate that lorcaserin , and likely other selective 5 - HT ( 2C ) receptor agonists , similarly affect both food - and nicotine-motivated behaviors , and nicotine-induced impulsivity . Collectively , these findings highlight a therapeutic potential for 5 - HT ( 2C ) agonists such as lorcaserin beyond obesity into addictive behaviors , such as nicotine dependence .

Gq-mediated Akt translocation to the membrane : a novel PIP 3 - independent mechanism in platelets . Akt is an important signaling molecule regulating platelet aggregation . Akt is phosphorylated after translocation to the membrane through Gi signaling pathways by a phosphatidylinositol -3,4 , 5 - trisphosphate ( PIP 3 ) - dependent mechanism . However , Akt is more robustly phosphorylated by thrombin compared with adenosine 5 ' - diphosphate in platelets . This study investigated the mechanisms of Akt translocation as a possible explanation for this difference . Stimulation of washed human platelets with protease-activated receptor agonists caused translocation of Akt to the membrane rapidly , whereas phosphorylation occurred later . The translocation of Akt was abolished in the presence of a Gq-selective inhibitor or in Gq-deficient murine platelets , indicating that Akt translocation is regulated downstream of Gq pathways . Interestingly , phosphatidylinositol 3 - kinase ( PI3K ) inhibitors or Q9H244 antagonist abolished Akt phosphorylation without affecting Akt translocation to the membrane , suggesting that Akt translocation occurs through a PI3K / PIP 3 / Gi-independent mechanism . An Akt scaffolding protein , P38936 - activated kinase ( PAK ) , translocates to the membrane after stimulation with protease-activated receptor agonists in a Gq-dependent manner , with the kinetics of translocation similar to that of Akt . Coimmunoprecipitation studies showed constitutive association of PAK and Akt , suggesting a possible role of PAK in Akt translocation . These results show , for the first time , an important role of the Gq pathway in mediating Akt translocation to the membrane in a novel Gi / PI3K / PIP 3 - independent mechanism .

P35367 occupancy in human brains after single oral doses of histamine H1 antagonists measured by positron emission tomography . 1 . P35367 occupancy in the human brain was measured in 20 healthy young men by positron emission tomography ( PET ) using [ 11C ] - doxepin . 2 . ( + ) - DB01114 MEN , a selective and classical antihistamine , occupied 76.8 + / - 4.2 % of the averaged values of available histamine H1 receptors in the frontal cortex after its administration in a single oral dose of 2 mg . Intravenous administration of 5 mg ( + ) - chlorpheniramine almost completely abolished the binding of [ 11C ] - doxepin to H1 receptors ( H1 receptor occupancy : 98.2 + / - 1.2 % ) . 3 . Terfenadine , a nonsedative antihistamine , occupied 17.2 + / - 14.2 % of the available H1 receptors in the human frontal cortex after its administration in a single oral dose of 60 mg . 4 . There was no correlation between H1 receptor occupancy by terfenadine and the plasma concentration of the active acid metabolite of terfenadine in each subject . 5 . PET data on human brain were essentially compatible with those on H1 receptor occupancy in guinea-pig brain determined by in vivo binding techniques , although for the same H1 receptor occupancy the dose was less in human subjects than in guinea-pigs . 6 . The PET studies demonstrated the usefulness of measuring H1 receptor occupancy with classical and second-generation antihistamines in human brain to estimate their unwanted side effects such as sedation and drowsiness quantitatively .

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 ( 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 , phospho - Q13158 , p16 ( INKA ) , Bax , Bak , and P38936 ( P38936 ) , 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 .

Modulation of a Mr 175,000 c-neu receptor isoform in Q9UBA6 / P00374 cells by serum starvation . The neu proto-oncogene product has been found to exist in two interconvertible forms in Q9UBA6 / P00374 mouse fibroblasts . The 185 - kilodalton form ( p185 ) present in growing cells is replaced by a 175 - kilodalton form ( p175 ) under conditions of serum starvation . This low molecular weight form accounts almost exclusively for the phosphotyrosine content of the receptor and is associated with increased tyrosine kinase activity . Addition of serum , platelet-derived growth factor or tumor promoter induces conversion of p175 to p185 within minutes , and this increase in molecular weight is associated with phosphorylation of serine and threonine ; removal of serum growth factors is followed by replacement of p185 with p175 over several hours . Unlike Q9UBA6 / P00374 cells , the human breast cancer cell line SK-Br - 3 expresses a high molecular weight neu / P04626 receptor with unchanged phosphotyrosine content in both serum-starved and serum-stimulated cultures . These findings indicate that activation of the neu proto-oncogene product in Q9UBA6 / P00374 cells may be regulated in part by protein kinase C-mediated receptor transmodulation rather than by ligand availability alone .

PPARgamma activation abolishes LDL-induced proliferation of human aortic smooth muscle cells via SOD-mediated down-regulation of superoxide . Native LDL would be a mitogenic and chemotactic stimulus of VSMC proliferation and differentiation in the atherosclerotic lesion where endothelial disruption occurred . In previous studies , our group investigated the molecular mechanisms by which LDL induces P10145 production and by which PPARalpha activation abolishes LDL effects in human aortic SMCs ( hAoSMCs ) . Herein is the first report of PPARgamma activation by troglitazone ( TG ) exerting its inhibitory effects on LDL-induced cell proliferation via generation not of H ( 2 ) O ( 2 ) , but of O2 ( . - ) , and the subsequent activation of Erk 1/2 in hAoSMCs . Moreover , in this study TG abolished the LDL-accelerated G ( 1 ) - S progression to control levels via down-regulation of active cyclinD 1 / P11802 and cyclinE / P24941 complexes and up-regulation of P38936 ( Cip 1 ) expression . TG exerted its anti-proliferative effects through the up-regulation of basal superoxide dismutase ( SOD ) expression . This data suggests that the regulation of O2 ( . - ) is located at the crossroads between LDL signaling and cell proliferation .

Systems pharmacology assessment of the 5 - fluorouracil pathway . AIM : To assess the impact of the 5 - fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC ( 50 ) for shRNA-exposed cells were three standard deviations outside the mean IC ( 50 ) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples .