interacts_with DB00834?
Molecular association between ATR and two components of the nucleosome remodeling and deacetylating complex , Q92769 and Q14839 . Q13315 ( Q13315 ) - and Rad3-related protein ( ATR ) is a phosphatidylinositol-kinase ( PIK ) -related kinase that has been implicated in the response of human cells to multiple forms of DNA damage and may play a role in the DNA replication checkpoint . The purification of an ATR complex allowed identification of chromodomain-helicase-DNA-binding protein 4 ( Q14839 ) as an ATR-associated protein by tandem mass spectrometric sequencing . Q14839 ( also called Mi-2beta ) is a component of a histone-deacetylase-2 ( Q92769 ) -containing complex , the nucleosome remodeling and deacetylating ( NRD ) complex . Endogenous ATR , Q14839 , and Q92769 are shown to coimmunoprecipitate , and ATR and Q92769 coelute through two biochemical purification steps . Other members of the NRD complex , Q13547 , Q13330 , and O94776 , are also detectable in ATR immunoprecipitates . ATR 's association with Q14839 and Q92769 suggests that there may be a linkage between ATR 's role in mediating checkpoints induced by DNA damage and chromatin modulation via remodeling and deacetylation . PEGylation of growth hormone-releasing hormone ( P01286 ) analogues . Synthetically produced GRF1-29 ( DB00010 ) has an amino acid composition identical to the N-terminal 29 amino acids sequence of the natural hypothalamic GHRH1-44 ( Figure 1 ) . It maintains bioactivity in vitro and is almost equally effective in eliciting secretion of endogenous growth hormone in vivo . The main drawbacks associated with the pharmaceutical use of hGRF1-29 relate to its short half-life in plasma , about 10-20 min in humans , which is caused mostly by renal ultrafiltration and enzymatic degradation at the N terminus . PEGylation has been considered as one valid approach to obtain more stable forms of the peptide , with a longer in vivo half-life and ultimately with increased pharmacodynamic response along the somatotropic axis ( endogenous GH , DB01277 levels ) . Different PEGylated P01286 conjugates were obtained and their bioactivity was tested in vitro and in vivo by monitoring endogenous growth hormone ( GH ) serum levels after intravenous ( i.v. ) injection in rats , and intravenous and subcutaneous ( s.c. ) injection in pigs . It was found that P01286 -PEG conjugates are able to bind and activate the human Q02643 , although with different potency . The effect of PEG molecular weight , number of PEG chains bound and position of PEGylation site on P01286 activity were investigated . Mono-PEGylated isomers with a PEG5000 polymer chain linked to Lys 12 or Lys 21 residues , showed high biological activity in vitro , which is similar to that of hGRF1-29 , and a higher pharmacodynamic response as compared to unmodified P01286 molecule . Possible involvement of endocannabinoids in the increase of morphine consumption in maternally deprived rat . Whether adolescent exposure to chronic DB00470 ( THC ) facilitates progression to opioid consumption is still controversial . In a maternal deprivation model ( 3 h daily from postnatal day 1-14 ) , we previously reported that adolescent exposure to chronic THC blocks morphine dependence in maternally deprived ( D ) rats . Owing to the existence of a functional cross-interaction between the opioid and cannabinoid systems in reward , we evaluated if the vulnerability to opiate reward in D rats , may involve an alteration of the endocannabinoid system . Anandamide and 2-arachidonoylglycerol ( 2-AG ) , were quantified in the striatum and mesencephalon of adolescent and adult D and non-deprived ( animal facility rearing , AFR ) rats by isotope dilution liquid chromatography-mass spectrometry . Oral morphine self-administration behavior was analyzed for 14 weeks , 24 days after chronic injection of the cannabinoid P21554 receptor antagonist/inverse agonist , SR141716A ( 3 mg/kg ) for 2 weeks during adolescence ( P01160 35-48 ) . Adolescent D rats exhibited higher basal levels of anandamide than adolescent AFR rats in the nucleus accumbens ( 38 % ) , the caudate-putamen nucleus ( 62 % ) and the mesencephalon ( 320 % ) , whereas adult D rats showed an increase of anandamide and 2-AG levels in the nucleus accumbens ( 50 % and 24 % , respectively ) and of 2-AG in the caudate-putamen nucleus ( 48 % ) , compared to adult AFR rats . Chronic administration of SR141716A to adolescent D rats blocked the escalation behavior in the morphine consumption test . Our data suggest that altered brain endocannabinoid levels may contribute to the escalation behavior in the morphine consumption test in a maternal deprivation model . P06401 -B regulation of insulin-like growth factor-stimulated cell migration in breast cancer cells via insulin receptor substrate-2 . Progesterone action contributes to the signaling of many growth factor pathways relevant to breast cancer tumor biology , including the insulin-like growth factor ( IGF ) system . Previous work has shown that insulin receptor substrate-2 ( Q9Y4H2 ) but not P35568 levels were regulated by progestin in progesterone receptor-B ( PR-B ) isoform expressing MCF-7 cells ( C4-12 PR-B ) . Furthermore , type 1 IGF receptor ( P08069 ) signaling via Q9Y4H2 correlated with the increased cell migration observed in a number of breast cancer cell lines . Consequently , in this study , we examined whether the elevation of Q9Y4H2 protein induced by progestin was sufficient to promote P05019 -stimulated cell motility . Treatment of C4-12 PR-B cells with progestin shifted the balance of phosphorylation from P35568 to Q9Y4H2 in response to P05019 . This shift in Q9Y4H2 activation was associated with enhanced migration in C4-12 PR-B cells pretreated with progestin , but had no effect on cell proliferation or survival . Treatment of C4-12 PR-B cells with DB00834 , an antiprogestin , inhibited IGF-induced cell migration . Attenuation of Q9Y4H2 expression using small interfering RNA resulted in decreased IGF-stimulated motility . In addition , Q9Y4H2 knockdown resulted in an abrogation of P31749 /Akt phosphorylation but not mitogen-activated protein kinase . Consequently , LY294002 , a phosphoinositide-3-kinase inhibitor , abolished IGF-induced cell motility in progestin-treated C4-12 PR-B cells . These data show a role for the PR in functionally promoting growth factor signaling , showing that levels of P41252 proteins can determine IGF-mediated biology , PR-B signaling regulates Q9Y4H2 expression , and that Q9Y4H2 can mediate IGF-induced cell migration via phosphoinositide-3-kinase in breast cancer cells . Analysis of common gene expression patterns in four human tumor cell lines exposed to camptothecin using cDNA microarray : identification of topoisomerase-mediated DNA damage response pathways . DB04690 ( CPT ) is a potent inhibitor of P11387 with a wide spectrum of anti-tumor activity . Relatively little information is available regarding the relation of known topoisomerase-mediated DNA damage with other intracellular pathways . To gain an insight into the intracellular molecular mechanisms of Topoisomerase I inhibitor camptothecin-mediated DNA damage leading to cell death , we used a high-density cDNA microarray to assess sensitive early gene expression profiles in SGC7901 ( gastric cancer ) , Hela ( cervical adenocarcinoma ) , K562 ( chronic myelogenous leukemia ) and HL60 ( promyelocytic leukemia ) tumor cells stimulated with camptothecin for 1 h at the concentrations of GI50 ( 50 % growth inhibition after 24 h of treatment ) . Analysis of the differentially expressed genes obtained 29 response genes common to all four cell lines . Moreover , these cell lines also shared the direction of regulation . Most of these common response genes were functionally related to cell proliferation or apoptosis , and some of them were involved in Q13315 ( ataxia-telangiectasia mutated ) and ATR ( Q13315 -and Rad3 related ) checkpoint pathways , JNK ( c-Jun N-terminal kinase ) pathway , the survival phosphatidylinositol ( PI ) 3 kinase-Akt-dependent pathway , mitochondrial cell death pathway , endoplasmic reticulum ( ER ) -related cell death pathway , and to ubiquitin/proteasome dependent protein degradation pathway . The data provides evidence for a linkage between topoisomerase-mediated DNA damage and intracellular signaling events , which may facilitate our understanding of the camptothecin mediated molecular mechanisms of action . Unsaturated side chain beta-11-hydroxyhexahydrocannabinol analogs . The cannabinoid side chain is a key pharmacophore in the interaction of cannabinoids with their receptors ( P21554 and CB2 ) . To study the stereochemical requirements of the side chain , we synthesized a series of cannabinoids in which rotation around the C1'- P06681 ' bond is blocked . The key steps in the synthesis were the cuprate addition of a substituted resorcinol to (+)-apoverbenone , the TMSOTf-mediated formation of the dihydropyran ring , and the stereospecific introduction of the beta-11-hydroxymethyl group . All the analogs tested showed nanomolar affinity for the receptors , the cis-hept-1-ene side chain having the highest affinity for P21554 ( Ki = 0.89 nM ) and showing the widest separation between P21554 and CB2 affinities . The parent n-heptyl-beta-11-hydroxyhexahydrocannabinol was the least potent binding to P21554 ( Ki = 8.9 nM ) and had the lowest selectivity between P21554 and CB2 . Q13639 receptor agonists increase sAPPalpha levels in the cortex and hippocampus of male C57BL/6j mice . BACKGROUND AND PURPOSE : A strategy to treat Alzheimer 's disease ( AD ) is to increase the soluble form of amyloid precursor protein ( sAPPalpha ) , a promnesic protein , in the brain . Because strong evidence supports beneficial effects of 5-hydroxytryptamine 5-HT(4) receptor agonists in memory and learning , we investigated the role of 5-HT(4) receptors on P05067 processing in 8 weeks-old male C57BL/6j mice . EXPERIMENTAL APPROACH : Mice were given , subcutaneously , prucalopride or ML 10302 ( s.c. ) , two highly selective 5-HT(4) receptor agonists and , up to 240 min later , the hippocampus and cortex were analysed by Western blot for sAPPalpha determination . KEY RESULTS : DB06480 ( 5 or 10 mg kg(-1) ) significantly increased sAPPalpha levels in the hippocampus and cortex , but did not modify the expression level of P05067 mRNA as detected by quantitative RT-PCR . A selective 5-HT(4) receptor antagonist , GR125487 ( 1 mg kg(-1) , s.c. ) inhibited prucalopride induced- increase in sAPPalpha levels . In addition , levels of sAPPalpha were increased by ML10302 only at 20 mg kg(-1) and was limited to the cortex . Also , prucalopride increased sAPPalpha levels in the cortex of a transgenic mouse model of AD , expressing the London mutation of P05067 . Furthermore , the combined injection of a selective acetylcholinesterase inhibitor , donepezil and prucalopride induced a synergic increase in sAPPalpha levels in the cortex and hippocampus . CONCLUSIONS AND IMPLICATIONS : Our results demonstrate that the 5-HT(4) receptor plays a key role in the non-amyloidogenic pathway of P05067 metabolism in vivo and give support to the beneficial use of 5-HT(4) agonists for AD treatment . Pharmacokinetic profiles of the novel P35354 selective inhibitor cimicoxib in dogs . DB05095 ( CX ) is a novel imidazole derivative that is a cyclo-oxygenase ( P36551 ) -2 selective non-steroidal anti-inflammatory drug and the latest P35354 selective inhibitor to be released for veterinary use . Currently there is limited information available on the pharmacokinetic ( PK ) properties of CX . The aim of the current study was to evaluate the PK features of CX after administration of the recommended dose and after administration of a more variable dose rate in the form of the commercially available tablet . In addition , the effects of food intake on the PK properties were also evaluated . In the first study , five healthy Beagle dogs received 2mg/kg CX via the oral route following a period of fasting . The second study was conducted using six healthy Labrador retriever dogs which each received an 80 mg tablet ( approximate dose 1.95-2.5mg/kg ) using a crossover design , both in the fasted and fed condition . The plasma concentrations of CX were detected by a validated HPLC method . No adverse effects were observed in any dogs during the experiment . The results from the PK analysis were similar between the studies , regardless of precision of dose and fasted and fed conditions . The mean peak concentration of CX was 0.49 and 0.43 μg/mL under fasted and fed conditions , respectively . The mean half-life was about 3h after all treatments . In addition , simulated multiple dosing data revealed that time over minimal effective concentration was similar after 1.95 , 2.0 and 2.5mg/kg dose administrations . These findings suggest that slight variation from the recommended dose should not alter the therapeutic outcome . In addition , CX can be administered to fed dogs without significantly affecting blood levels . P01133 -like ligands mediate progesterone 's anti-apoptotic action on macaque granulosa cells . A local autocrine/paracrine role for progesterone is an absolute requirement for corpus luteum formation in primates . Despite this , the mechanism(s) remain obscure , although existing data suggest an anti-apoptotic action to be central . There are a limited number of progestin-regulated gene targets identified in the luteinizing primate follicle , suggesting that a small number of important genes may mediate progesterone action . Possible gene targets could be the epidermal growth factor ( P01133 ) family members amphiregulin ( P15514 ) and epiregulin ( O14944 ) . Using macaques undergoing controlled ovarian stimulation cycles , we show that the phosphorylation of P01133 receptor ( P00533 ) , P29323 1/2 , and AKT increases 6 h after an ovulatory human chorionic gonadotropin ( hCG ) stimulus and remains activate through 24 h . Immunoreactive O14944 and P15514 ligands in the follicular fluid both increased in a time frame commensurate with P00533 phosphorylation . The mRNA expression of P15514 and O14944 in nonluteinized granulosa cells ( NLGC ) was induced in culture with hCG , an effect blocked by progesterone receptor ( P06401 ) antagonists . Overexpression of P06401 B in NLGC and treatment with a nonmetabolizable progestin did not increase either gene , indicating both progesterone and luteinizing hormone/CG are necessary . Addition of P01133 and P01133 -like ligands did not promote steroidogenesis in vitro by granulosa cells in the presence of gonadotropin , but were able to partially reverse DB00834 -induced cell death . These data suggest that progesterone promotes the expression of P15514 and O14944 , which in turn maintain viability of luteinizing granulosa cells , representing one possible mechanism whereby progesterone promotes corpus luteum formation in the primate . DB00583 -palmitoyltransferase 2 deficiency : novel mutations and relevance of newborn screening . We report on a newborn male , born at term after an uneventful pregnancy presenting with a pathological acylcarnitine profile in routine newborn screening on the third day of life . The profile showed characteristic elevations of C14:0- , C16:0- , C16:1- and C18:1-acylcarnitines , while the ratio of ( C16 + C18:1 ) / P06681 was increased , suggesting P23786 - or carnitine-acylcarnitine-translocase- deficiency . The acylcarnitine profile in blood taken on day 9 was normal with breast milk feeding . No dicarboxylic aciduria was found . In fibroblasts , the activity of P23786 was decreased to 25 % , overall oxidation of the long-chain fatty acids was reduced to 10 % of control values . Sequence analysis of the P23786 gene showed heterozygosity for two previously undescribed mutations in exon 4 : c.748-749delAA ( truncating ) , and c.1436A > G ( p.Tyr479Cys ; missense ) mutations . The asymptomatic parents were found to be heterozygous , the mother carries the c.748-749delAA and the father the c.1436A > G mutation . The boy is now 2.5 years old ; no clinical symptoms associated with the marked impairment of long-chain fatty acid oxidation have occurred . Confirmation of mitochondrial fatty acid oxidation defects from an initial abnormal newborn-screening by tandem mass spectrometry should include enzyme and , if possible , molecular genetic analysis despite a normal 2nd screening . Biochemical testing of urine ( organic acids ) may be unrevealing . No effect of the oral neutral endopeptidase inhibitor candoxatril , on bronchomotor tone and histamine reactivity in asthma . P08473 ( NEP ) is found in many tissues in man , including the lung . Metabolism by NEP is one of the main mechanisms for the clearance of atrial natriuretic peptide ( P01160 ) , a hormone that causes bronchodilation and reduces nonspecific bronchial reactivity in man . DB00616 , an oral NEP inhibitor has been shown to elevate circulating P01160 levels . We have sought to determine whether the administration of candoxatril will alter bronchomotor tone ( forced expiratory volume in one second ( FEV1 ) ) and histamine reactivity . Ten male asthmatic patients with stable asthma were enrolled ( mean ( SD ) age 32 ( 10 ) yrs ; FEV1 92 ( 11 ) % predicted ) in a randomized , double-blind , placebo-controlled study . On each study day , after baseline spirometry , patients received 200 mg of candoxatril or placebo . Spirometry was repeated at half hourly intervals . After 2 h a histamine inhalation test was performed . There was no significant difference in FEV1 values at baseline or at 2 h post-dosing between active and placebo study days , with mean ( SEM ) FEV1 at baseline and 2 h of 3.71 ( 0.29 ) l and 3.85 ( 0.29 ) l on the placebo day , and 3.89 ( 0.27 ) l and 4.05 ( 0.82 ) l on the active day , respectively . The geometric mean ( range ) provocative concentration of histamine producing a 20 % fall in FEV1 ( PC20 ) on the placebo day and active day did not differ significantly , being 1.17 ( 0.25-25.8 ) and 0.93 ( 0.13-32 ) mg.ml-1 , respectively. ( ABSTRACT TRUNCATED AT 250 WORDS ) Antiinflammatory steroid action in human ovarian surface epithelial cells . The human ovarian surface epithelium ( OSE ) is subject to serial injury and repair during ovulation , which is a natural inflammatory event . We asked whether there is a compensatory antiinflammatory component to this process , involving steroid hormones produced locally at the time of ovulation . Quantitative RT-PCR analysis of total RNA from cultured human OSE cell monolayers showed that exposure to proinflammatory IL1alpha ( 500 pg/ml ) increased mRNA levels of cyclooxygenase-2 ( P35354 ) ( P < 0.01 ) at 48 h . The P35354 mRNA response to IL1alpha was associated with an approximate 18-fold ( P < 0.01 ) increase in mRNA levels of 11beta-hydroxysteroid dehydrogenase type 1 ( 11betaHSD1 ) , encoding the steroid dehydrogenase that reversibly reduces cortisone to antiinflammatory cortisol . Addition of cortisol to OSE cell culture medium dose-dependently suppressed the P35354 mRNA response to IL1alpha ( P < 0.01 ) but reciprocally enhanced the 11betaHSD1 mRNA response ( P < 0.05 ) , with both effects strongest at 1 microm cortisol . Presence of glucocorticoid receptor-alpha mRNA and protein was established in OSE cell monolayers and treatment with IL1alpha shown to significantly up-regulate the glucocorticoid receptor-alpha mRNA level ( P < 0.05 ) . P04150 antagonist ( DB00834 , 10 microm ) fully reversed the inhibitory effect of 1 microm cortisol on IL1alpha-stimulated P35354 mRNA expression . Progesterone also suppressed IL1alpha-induced P35354 mRNA expression but had no significant effect on IL1alpha-stimulated 11betaHSD1 expression . These data provide direct evidence for antiinflammatory actions of cortisol and progesterone in human OSE cells . Distribution patterns of cannabinoid P21554 receptors in the hippocampus of APPswe/PS1ΔE9 double transgenic mice . Cannabinoids have neuroprotective effects that are exerted primarily through cannabinoid P21554 receptors in the brain . This study characterized P21554 receptor distribution in the double transgenic ( dtg ) P05067 (swe)/ P49768 (ΔE9) mouse model for Alzheimer 's disease . Immunohistochemical labeling of P21554 protein in non-transgenic mice revealed that P21554 was highly expressed in the hippocampus , with the greatest density of P21554 protein observed in the combined hippocampal subregions P00918 and P07451 ( P00918 /3 ) . P21554 immunoreactivity in the P00915 and P00918 /3 hippocampal regions was significantly decreased in the dtg P05067 (swe)/ P49768 (ΔE9) mice compared to non-transgenic littermates . Reduced P21554 expression in dtg P05067 (swe)/ P49768 (ΔE9) mice was associated with astroglial proliferation and elevated expression of the cytokines inducible nitric oxide synthase and tumor necrosis factor alpha . This finding suggests an anti-inflammatory effect of cannabinoids that is mediated by P21554 receptor , particularly in the P00918 /3 region of the hippocampus . Furthermore , the study suggests a decreased P21554 receptor expression may result in diminished anti-inflammatory processes , exacerbating the neuropathology associated with Alzheimer 's disease . Targeted cancer therapies based on antibodies directed against epidermal growth factor receptor : status and perspectives . Compelling experimental and clinical evidence suggests that epidermal growth factor receptor ( P00533 ) plays an important role in the pathogenesis of a variety of human cancers ; thus , providing a strong rationale for the development of receptor antagonists as effective and specific therapeutic strategies for the treatment of P00533 -expressing cancers . Monoclonal antibodies ( mAb ) , owing to their high specificity towards a given target , represent a unique class of novel cancer therapeutics . A number of anti- P00533 mAb are currently being developed in our clinic , including two that have been approved by the United States Food and Drug Administration for the treatment of refractory metastatic colorectal cancer ( mCRC ) and squamous cell carcinomas of the head and neck ( SCCHN ) . Cetuximab ( Erbitux , IMC-C225 ) , an IgG1 mAb , has demonstrated significant antitumor activity , both as a single agent and in combination with chemotherapeutics and radiation , in patients with refractory mCRC and SCCHN , respectively . DB01269 ( Vectibix ) , an IgG2 mAb , has been approved as a single agent for the treatment of patients with refractory mCRC . These mAb , via blocking ligand/receptor interactions , exert their biological activity via multiple mechanisms , including inhibition of cell cycle progression , potentiation of cell apoptosis , inhibition of DNA repair , inhibition of angiogenesis , tumor cell invasion and metastasis and , potentially , induction of immunological effector mechanisms . Anti- P00533 antibodies have demonstrated good safety profiles and potent anticancer activity in our clinic and may prove to be efficacious agents in the treatment of a variety of human malignancies . P04150 antagonism disrupts the reconsolidation of social reward-related memories in rats . Reconsolidation is the process whereby consolidated memories are destabilized upon retrieval and restabilized to persist for later use . Although the neurobiology of the reconsolidation of both appetitive and aversive memories has been intensively investigated , reconsolidation of memories of physiologically relevant social rewards has received little attention . Social play , the most characteristic social behaviour displayed by young mammals , is highly rewarding , illustrated by the fact that it can induce conditioned place preference ( CPP ) . Here , we investigated the role of signalling mechanisms implicated in memory processes , including reconsolidation , namely glucocorticoid , mineralocorticoid , DB01221 glutamatergic and P21554 cannabinoid receptors , in the reconsolidation of social play-induced CPP in rats . Systemic treatment with the glucocorticoid receptor antagonist mifepristone before , but not immediately after , retrieval disrupted the reconsolidation of social play-induced CPP . DB00834 did not affect social play-induced CPP in the absence of memory retrieval . Treatment with the DB01221 receptor antagonist MK-801 modestly affected the reconsolidation of social play-induced CPP . However , the reconsolidation of social play-induced CPP was not affected by treatment with the mineralocorticoid and P21554 cannabinoid receptor antagonists spironolactone and rimonabant , respectively . We conclude that glucocorticoid neurotransmission mediates the reconsolidation of social reward-related memories in rats . These data indicate that the neural mechanisms of the reconsolidation of social reward-related memories only partially overlap with those underlying the reconsolidation of other reward-related memories . Peripheral endocannabinoid system-mediated actions of rimonabant on growth hormone secretion are ghrelin-dependent . The somatotroph axis is a crucial pathway regulating metabolism . Despite the fact that the endocannabinoid system has been also revealed as a potent modulator of energy homeostasis , little information is available concerning a putative interaction between these two systems . The aim of the present study was to determine the in vivo effects of the blockade of the cannabinoid receptor type 1 ( P21554 ) over growth hormone ( GH ) secretion using the P21554 antagonist rimonabant . The results obtained show that the blockade of the P21554 peripheral receptor by i.p. injection of rimonabant significantly inhibited pulsatile GH secretion . Similarly , it was found that this injection significantly decreased ghrelin-induced GH secretion without any effect on growth hormone-releasing hormone ( P01286 ) -induced GH discharge . In situ hybridisation showed that the peripheral blockade of P21554 did not affect hypothalamic somatostatin mRNA levels ; however , P01286 mRNA expression was significantly decreased . The blockade of the vagus nerve signal by surgical vagotomy eliminated the inhibitory action of rimonabant on P01286 mRNA and consequently on GH . On the other hand , the central P21554 blockade by i.c.v. rimonabant treatment was unable to reproduce the effect of peripheral blockade on P01286 mRNA , nor the GH response to ghrelin . In conclusion , the data reported in the present study establish , from a physiological point of view , the existence of a novel mechanism of GH regulation implicating the action of the cannabinoid receptor on the somatotroph axis . P04150 and histone deacetylase-2 mediate dexamethasone-induced repression of P98088 gene expression . Airway occlusion in obstructive airway diseases is caused in part by the overproduction of secretory mucin glycoproteins through the up-regulation of mucin ( MUC ) genes by inflammatory mediators . Some pharmacological agents , including the glucocorticoid dexamethasone ( DB00514 ) , repress mucin concentrations in lung epithelial cancer cells . Here , we show that DB00514 reduces the expression of P98088 , a major airway mucin gene , in primary differentiated normal human bronchial epithelial ( NHBE ) cells in a dose-dependent and time-dependent manner , and that the DB00514 -induced repression is mediated by the glucocorticoid receptor ( GR ) and two glucocorticoid response elements ( GREs ) in the P98088 promoter . The pre-exposure of cells to DB00834 , a GR antagonist , and mutations in either the GRE3 or GRE5 cis-sites abolished the DB00514 -induced repression . Chromatin immunoprecipitation ( ChIP ) assays showed a rapid temporal recruitment of GR to the GRE3 and GRE5 cis-elements in the P98088 promoter in NHBE and in A549 cells . Immunofluorescence showed nuclear colocalization of GR and histone deacetylase-2 ( Q92769 ) in P98088 -expressing NHBE cells . ChIP also showed a rapid temporal recruitment of Q92769 to the GRE3 and GRE5 cis-elements in the P98088 promoter in both cell types . The knockdown of Q92769 by Q92769 -specific short interfering RNA prevented the DB00514 -induced repression of P98088 in NHBE and A549 cells . These data demonstrate that GR and Q92769 are recruited to the GRE3 and GRE5 cis-sites in the P98088 promoter and mediate the DB00514 -induced cis repression of P98088 gene expression . A better understanding of the mechanisms whereby glucocorticoids repress P98088 gene expression may be useful in formulating therapeutic interventions in chronic lung diseases . AM251 induces apoptosis and G2/M cell cycle arrest in A375 human melanoma cells . Human cutaneous melanoma is an aggressive and chemotherapy-resistant type of cancer . AM251 is a cannabinoid type 1 ( P21554 ) receptor antagonist/inverse agonist with off-target antitumor activity against pancreatic and colon cancer cells . The current study aimed to characterize the in-vitro antimelanoma activity of AM251 . The P15056 V600E mutant melanoma cell line , A375 , was used as an in-vitro model system . Characterization tools included a cell viability assay , nuclear morphology assessment , gene expression , western blot , flow cytometry with P08758 -FITC/7-AAD double staining , cell cycle analyses , and measurements of changes in intracellular DB02527 and calcium concentrations . AM251 exerted a marked cytotoxic effect against A375 human melanoma cells with potency comparable with that observed for cisplatin without significant changes in the human dermal fibroblasts viability . AM251 , at a concentration that approximates the IC50 , downregulated genes encoding antiapoptotic proteins ( P10415 and survivin ) and increased transcription levels of proapoptotic Q07812 , induced alteration of P08758 reactivity , DNA fragmentation , chromatin condensation in the cell nuclei , and G2/M phase arrest.AM251 also induced a 40 % increase in the basal DB02527 levels , but it did not affect intracellular calcium concentrations . The involvement of Q9Y2T6 , O75762 , and P35354 in the AM251 mechanism of action was excluded . The combination of AM251 with celecoxib produced a synergistic antitumor activity , although the mechanism underlying this effect remains to be elucidated . This study provides the first evidence of a proapoptotic effect and G2/M cell cycle arrest of AM251 on A375 cells . This compound may be a potential prototype for the development of promising diarylpyrazole derivatives to be evaluated in human cutaneous melanoma . Factors regulating insulin-like growth factor-binding protein-3 binding , processing , and potentiation of insulin-like growth factor action . In this study , we investigated the effects of various biochemical and pharmacological agents on insulin-like growth factor ( IGF ) -binding protein-3 ( P17936 ) cell binding and action in cultured bovine fibroblasts . When cells were preincubated for 48 h with 50 nM recombinant human ( rh ) P17936 , P05019 -stimulated [3H]aminoisobutyric acid ( [125H]AIB ) uptake was enhanced 2- to 3-fold . The addition of cytoskeletal disrupting agents during the preincubation with DB05897 did not affect P17936 potentiation of P05019 action , nor did a variety of serine , aspartate , and metalloproteinase inhibitors . On the other hand , ammonium chloride and chloroquine , weak bases that neutralize the pH of acidic cell compartments , blocked P17936 potentiation of P05019 -stimulated [3H]AIB uptake . Chloroquine and ammonium chloride had no effect alone and did not inhibit P08069 binding or action in the absence of DB05897 . Bafilomycin A , a specific inhibitor of DB00171 -dependent hydrogen ion pumps , also inhibited P17936 potentiation of P05019 -stimulated [3H]AIB uptake . Competitive [125I] P05019 binding and affinity cross-linking experiments suggested structure/function changes in cell-bound P17936 that were altered in the presence of chloroquine and bafilomycin . DB01109 markedly decreased initial P17936 cell adherence , but could not promote dissociation of P17936 from cells after the 48-h preincubation . Moreover , heparin did not inhibit P17936 potentiation of P05019 action . In summary , these data indicate that P17936 undergoes specific pH-dependent structural and/or environmental modifications that mediate the enhancing effect of P17936 on P05019 action in bovine fibroblasts . They also suggest that P17936 binding to heparin-like molecules on the cell surface is not directly involved in this process . DB08881 -induced interface dermatitis manifesting as radiation-recall and a keratosis pilaris-like eruption . DB08881 is a specific inhibitor of the V600E mutated P15056 protein kinase used for the treatment of unresectable or metastatic melanoma harboring this mutation . Multiple predictable side effects have been described with use of this targeted therapy , and implicate P15056 and mitogen activated protein kinase ( MAPK ) signaling pathways in their pathogenesis . Herein , we report the novel finding of an interface dermatitis in radiation recall and a keratosis pilaris-like clinical reaction in a patient treated with vemurafenib .