黄--干扰miR-21表达对子宫内膜癌顺铂耐药细胞株IshikawaDDP的影响.docx

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1、干扰miR-21表达对子宫内膜癌顺铂耐药细胞株Ishikawa/DDP的影响[摘要]目的观察干扰miR-21表达对子宫内膜癌顺铂耐药细胞株Ishikawa/DDP的影响。方法以Lipofectamine2000介导miR-21抑制剂转染Ishikawa/DDP细胞株，同时设置阴性组和耐药组。采用逆转录PCR检测miR-21、多药耐药基因MDR1、促凋亡基因Bax和抗凋亡基因Bcl-2的表达。采用蛋白印迹法检测多药耐药蛋白P-gp、促凋亡蛋白Bax和抗凋亡蛋白Bcl-2的表达。采用噻唑蓝比色法检测细胞对顺铂的敏感性。采用

2、流式细胞术检测细胞凋亡情况。结果转染miR-21抑制剂后，抑制剂组miR-21、MDR1、P-gp和Bcl-2的表达显著低于耐药组和阴性组（P＜0.05）；而抑制剂Bax的表达显著高于耐药组和阴性组（P＜0.05）；耐药组和阴性组各基因蛋白的表达无明显差异（P＞0.05）。顺铂对抑制剂组的IC50值显著低于耐药组和阴性组（P＜0.05）；顺铂对抑制剂组细胞的诱导凋亡率显著大于耐药组和阴性组（P＜0.05）；耐药组与阴性组的IC50值和细胞凋亡率比较无统计学差异（P＞0.05）。结论干扰miR-21表达可显著提高Ishi

3、kawa/DDP细胞株对顺铂的敏感性，并促进细胞凋亡，其具体机制可能与下调MDR1、P-gp和Bcl-2表达，以及上调Bax表达有关。[关键词]miR-21；子宫内膜癌；顺铂；耐药EffectofinterferingtheexpressionofmiR-21onIshikawa/DDPcisplatinresistantcellinendometrialcarcinoma[Abstract]ObjectiveToobservetheeffectofinterferingtheexpressionofmiR-21onI

4、shikawa/DDPcisplatinresistantcellinendometrialcarcinoma.MethodsIshikawa/DDPcisplatinresistantcellwastransfectedwithmiR-21inhibitorbyLipofectamine2000.Atthesametimesettingofthecontrolgroupandthedrugresistantgroup.TheexpressionofmiR-21,MDR1,BaxandBcl-2wasdetectedb

5、yreversetranscriptionPCR.TheexpressionofP-gp,BaxandBcl-2wasdetectedbyWesternblot.ThesensitivityofIshikawa/DDPcellstocisplatinwasdetectedbyMTT.Thecellapoptosiswasdetectedbyflowcytometry.ResultsAftertransfectionwithmiR-21inhibitor,theexpressionofmiR-21,MDR1,P-gpan

6、dBcl-2intheinhibitorgroupwassignificantlylowerthanthatinthedrugresistantgroupandthenegativegroup(P＜0.05).TheexpressionofBaxintheinhibitorgroupwassignificantlyhigherthanthatinthedrugresistantgroupandthenegativegroup(P＜0.05).Therewasnosignificantdifferenceintheexp

7、ressionofgeneandproteininthedrugresistantgroupandthenegativegroup(P＞0.05).TheIC50valueintheinhibitorgroupwassignificantlylowerthanthatinthedrugresistantgroupandthenegativegroup(P＜0.05).Theapoptosisrateintheinhibitorgroupwassignificantlyhigherthanthatinthedrugres

8、istantgroupandthenegativegroup(P＜0.05).TherewasnosignificantdifferenceinIC50andapoptosisratebetweenthetwogroups(P＞0.05).ConclusionInterferetheexpressionofmiR-21cansig