胡椒碱与棉酚联用对前列腺癌DU145细胞生长抑制的协同作用_曾龙辉

7/19/20121548http//．cnki．net/kcms/detail/34．1086．R．20120719．1548．015．htmlDU1451211111．2．5106322012－03－[e-mail][e-mail]10．3969/j．issn．1001－1978．2012．08．015A1001－1978201208－1101－05R284.1R329.28R737.250.22R979.1DU145。MTSCYP3A4。。IC5021.00μmol·L－18.07μmol·L－1CDI＜1。G0/G1p21Cip1CyclinD1、CyclinA、CyclinB1CYP3A4。CYP3A4。DU145CYP3A4gossypolGOS、。。BclBH3Bcl1。2－5、、、DNA。6。piperine、、、CYP3A4P4503A42000%154%7－8。DU145。11．1DU145DMEMfetalbovineserumFBS、L-、Gibco/Invitrogengossypolaceticacid、propidiumiodidePIdimethylsulfoxideDMSOSigma-AldrichpiperineMTSCell-Titer96AqueousONEsolutionCellProliferationAs-sayPromegaCyclinA、CyclinB1、CyclinD1、CyclinEp21Cip1CellSig-nalingTechnologyCYP3A4Milli-pore。1．21．2．1DU14510%、100kU·L－1100mg·L－1DMEM37℃、5%CO22～3d。1．2．2MTS3.30×107·L－196100μl2。48h20μlMTS37℃1～4h。490nm。GraphPadPrism4.0IC50。1．2．3100μmol·L－1、15μmol·L－1·1011·ChinesePharmacologicalBulletin2012Aug2881101～5100μmol·L－1+15μmol·L－1。24h48hPBS2－20℃。PI37℃1h。1．2．4DU1451.80×10675cm2。24h。6h24hPBS2×SDSSDS-PAGE。PVDF1h4℃。PBST31hBeyoECLPlusX-。1．2．5coefficientofdruginteractionCDICDICDI=AB/A×B。ABAB。CDI＜1CDI＜0.7CDI=1CDI＞1。Graph-PadPrism4.0Newman-Keuls。22．1DU145DU145IC50226.68μmol·L－1100μmol·L－1Fig1A。100μmol·L－1DU145。IC5021.00μmol·L－18.07μmol·L－1Fig1B。CDI100μmol·L－110～20μmol·L－1CDI0.7Fig1C。2．2G0/G124hG0/G1SG0/G1G2/Msub-G0/G1。48hG0/G1、SG2/MP＜0.01Fig2。Fig1SynergisticinhibitionoftheproliferationofDU145cellsbypiperineplusGOSAPiperineinhibitstheviabilityofDU145cellsinadose-dependentmannerBGOSshowsadose-dependentcytotoxicityinDU145cellsandwhenitiscombinedwith100μmol·L－1piperinethecytotoxicityincreasessignificantlyCCDI＜1indicatesthatpiperineplusgossypolhasasynergisticeffectoninhibitionofgrowthofDU145cells．·2011·ChinesePharmacologicalBulletin2012Aug288Fig2Effectsofpiperineand/orGOSoncellcycledistributionofDU145cells*P＜0.05＊＊P＜0.012．3cyclincyclin-dependentkinaseCDK。CyclinDCyclinEG1/SCyclinASDNAG2CyclinACyclinB。p21Cip1CDKp21Cip1CDK。Fig36hp21Cip1CyclinD1CyclinB1。24hp21Cip1CyclinD1、CyclinA、CyclinB1p21Cip1CyclinD1、CyclinA、CyclinB1。CyclinE。G0/G1。Fig3EffectsofpiperinecombinedwithGOSoncellcyclerelatedproteinsexpressionTherelativedensitometryvaluesofcellcyclerelatedproteinsagainsttheaverageoftotalβ-tubulinarelistedundereachofthebands．2．4CYP3A4CYP3A4P450。CYP3A4。Fig46h24hCYP3A4。6hCYP3A4CYP3A4。Fig4Effectsofpiperineand/orGOSontheexpressionofCYP3A4inDU145cellsTherelativedensitometryvaluesofCYP3A4againsttheaverageoftotalβ-tubulinareshowedundereachofthebands．39－10。DU145CDI＜1·3011·ChinesePharmacologicalBulletin2012Aug288。DU145G0/G1p21Cip1DNA。DU145p21Cip1DNA3－5。15μmol·L－1DU14548h。DU14524h48h。DNADU145。CYP3A411。CYP3A4P45040%50%12。CYP3A413。CYP3A4CYP3A4。CYP3A4DU145。CYP3A4BH3DU14514。70。。。。1MengYTangWHDaiYetal．NaturalBH3mimetic－-gossypolchemosensitizeshumanprostatecancerviaBcl-xLinhibi-tionaccompaniedbyincreaseofPumaandNoxaJ．MolCancerTher2008772192－202．2．JurkatTJ．2008306742－6．2XuWBXuLHLuHSetal．Effectsofgossypolontheprolif-erationandapoptosisofJurkatTcellsJ．ChinJCellBiol2008306742－6．3LiguerosMJeoungDTangBetal．Gossypolinhibitionofmito-siscyclinD1andRbproteininhumanmammarycancercellsandcyclin-D1transfectedhumanfibrosarcomacellsJ．BrJCancer199776121－8．4VanPoznakCSeidmanADReidenbergMMetal．Oralgos-sypolinthetreatmentofpatientswihrefractorymetastaticbreastcancerAphaseⅠ/ⅡclinicaltrialJ．BreastCancerRes2001663239－48．5ZhangMCLiuHPGuoRBetal．Molecularmechanismofgossypol-inducedcellgrowthinhibitionandcelldeathofHT-29humancoloncarcinomacellsJ．BiochemPharmacol200366193－103．6HaspelHCRenYFWatanabeKAetal．CytocidaleffectofgossypolonculturedmurineerythroleukemiacellsispreventedbyserumproteinJ．JPharmacolExpTher19842291218－25．7BhardwajRKGlaeserHBecquemontLetal．Piperineama-jorconstituentofblackpepperinhibitshumanP-glycoproteinandCYP3A4J．JPharmacolExpTher20023022645－50．8ShobaGJoyDJosephTetal．InfluenceofpiperineonthepharmacokineticsofcurcumininanimalsandhumanvolunteersJ．PlantMed1998644353－6．9SunilaESKuttanG．ImmunomodulatoryandantitumoractivityofPiperlongumLinn．andpiperineJ．JEthnopharmacol2004902－3339－46．10BezerraDPPessoaCdeMoraesMOetal．AntiproliferativeeffectsoftwoamidespiperineandpiplartinefromPiperspeciesJ．ZNaturforschC2005607－8539－43．11．CYP3A4J．201228116－9．11LiMZhuCJ．AdvancesinCYP3A4over-expressedcellmodelsandtheirapplicationtodrugmetabolismresearchJ．ChinPhar-macolBull201228116－9．12．CYP4503A4J．200925121619－22．12ZhangZHWuHHXueGetal．Naringenininhibitstheup-regulationofCYP3A4andattenuatesthecellulartoxicityinducedbyrifampicinandisoniazidinhepatocytesJ．ChinPharmacolBull200925121619－22．13PeterMKonstantinGDanielCetal．Co-administrationofpip-erineanddocetaxelresultsinimprovedanti-tumorefficacyviainhi-bitionofCYP3A4activityJ．Prostate2012726661－7．14OuyangDYJiYHMarkSetal．ValproicacidsynergisticallyenhancesthecytotoxicityofgossypolinDU145prostatecancercellsAniTRTAQ-basedquantitativeproteomicanalysisJ．JProteomics201174102180－93．·4011·ChinesePharmacologicalBulletin2012Aug288SynergisticinhibitionofgrowthofprostatecancerD