孕激素依赖的cyclin G1在小鼠子宫内膜上皮细胞的表达及其对细胞增殖的影响VIP专享VIP免费

生理学报ActaPhysiologicaSinica,August25,2008,60(4):541-546http://www.actaps.com.cn541研究论文Received2008-01-21Accepted2008-04-25ThisworkwassupportedbytheNationalNaturalScienceFoundationofChina(No.30770813).*Correspondingauthor.Tel:+86-28-85503048;Fax:+86-28-85503204;E-mail:yuelimin@yahoo.com孕激素诱导cyclinG1在小鼠子宫内膜上皮细胞的表达及其对细胞增殖的影响马琰岩，范轶，白玛康卓，张金虎，何亚萍，喻琳鳞，岳利民*四川大学华西基础医学与法医学院生理学教研室，成都610041摘要：本文在体外培养条件下研究卵巢激素诱导小鼠子宫内膜上皮细胞cyclinG1的表达及细胞增殖和细胞周期进程的变化，以探讨孕激素依赖的细胞周期调控因子cyclinG1对子宫内膜上皮细胞增殖的负调控作用。原代培养小鼠子宫内膜上皮细胞，待其生长汇合后分为4组：对照组(C组)、雌激素组(E组)、孕激素组(P组)、雌、孕激素共同作用组(EP组)。加入相应激素作用24h后，用细胞免疫化学方法检测各组细胞cyclinG1的表达水平；四甲基偶氮唑蓝(MTT)比色法检测各组细胞活力，间接观察子宫内膜上皮细胞的增殖情况；用流式细胞仪检测分布在细胞周期各时相的子宫内膜上皮细胞所占百分数。细胞免疫化学结果显示，cyclinG1在C组和E组子宫内膜上皮细胞上无明显表达，而在P组和EP组子宫内膜上皮细胞中表达明显，且定位于细胞核内。MTT法结果显示，与C组相比，E组细胞活力明显增高，而P组和EP组的细胞活力均明显下降，表明雌激素能促进子宫内膜上皮细胞增殖，而孕激素则具有抑制子宫内膜上皮细胞增殖的作用。流式细胞术检测显示，与C组相比，E组中处于S期的子宫内膜上皮细胞百分数增多；P组与EP组中处于S期的子宫内膜上皮细胞百分数明显减少，而处于G1期的细胞百分数和G2/M期的细胞百分数则明显增加。上述结果提示，孕激素依赖的cyclinG1可能通过阻滞细胞周期进程来参与孕激素对子宫内膜上皮细胞增殖的负调控作用。关键词：细胞周期素G1；子宫内膜；细胞周期；孕激素中图分类号：R339.2Theprogesterone-inducedexpressionofcyclinG1anditseffectontheproliferationofmouseuterineepithelialcellsMAYan-Yan,FANYi,BaiMa-Kang-Zhuo,ZHANGJin-Hu,HEYa-Ping,YULin-Lin,YUELi-Min*DepartmentofPhysiology,WestChinaSchoolofPreclinicalandForensicMedicine,SichuanUniversity,Chengdu610041,ChinaAbstract:Theaimofthepresentstudyistoinvestigatetheeffectofprogesterone-inducedexpressionofcyclinG1ontheproliferationofendometrialepithelialcells.Toobtainmouseendometrialepithelialcells,theuteriwereisolatedfromovariectomizedmicewhichwereinjectedsubcutaneouslywith100ngestradiolperdayfortwodays.Thentheuteriweredigestedbydispaseandpancreatinrespectively.EndometrialepithelialcellswereculturedinDMEM/F12containing6%fetalbovineserum,anddividedintofourgroupswhentheygrewtoconfluence.Eachofthegroupswastreatedasfollows:GroupEwastreatedwith0.01μmol/Lestradiolonly,groupPwastreatedwith1μmol/Lprogesterone,groupEPwastreatedwithboth0.01μmol/Lestradioland1μmol/Lprogesterone,andgroupCwastreatedwith0.01%DMSOforcontrol.ImmunocytochemistrywasusedtoexaminetheexpressionofcyclinG1protein.MTTassaywasusedtoevaluatemetabolicactivityofcells.Flowcytometrywasusedtocheckthenumberofcellsdistributingineachphaseofthecellcycle.TheresultofimmunocytochemistryshowedthattherewasnoexpressionofcyclinG1proteiningroupCandgroupE,whilecyclinG1wasobviouslyexpressedingroupPandgroupEPandlocalizedinnucleus.IntheMTTassay,comparedwithgroupC,theviabilityofgroupEsignificantlyincreased,whilethatofbothgroupPandgroupEPdecreasedsignificantly.TheresultsofflowcytometrywereinaccordancewiththoseofMTT,whichshowedthatcomparedwithgroupC,groupEhadahigherproportionofcellsinSphase,whilegroupP,aswellasgroupEPhadalowerproportionofcellsinSphasebutahi...