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1、·674·南方医科大学学报（JSouthMedUniv）2011;31(4)·基础研究·特异性miR-221抑制剂对结直肠癌细胞增殖及凋亡的影响王伟，孙凯，吴承堂，雷尚通，曾俊杰，伍颖君，李国新南方医科大学南方医院普通外科，广东广州510515摘要：目的探讨miR-221在结直肠癌细胞中的表达状况及特异性miR-221抑制剂对癌细胞增殖和凋亡的影响。方法应用实时荧光定量PCR（real-timeQ-PCR）检测四种人结肠癌细胞系HT-29、Lovo、SW-480、Caco2中miRNA-221表达状况，并以人脐静脉内皮细胞HUVEC作为正常对照；设计并合成miR-221、anti
2、-miR-221（微小RNA抑制剂）寡核苷酸，以脂质体转染Caco2细胞系，以Real-timeQ-PCR再次检测转染后细胞中miR-221表达状况，并通过噻唑蓝（MTT）比色法及流式细胞仪观察细胞的增殖和凋亡状态。结果与正常细胞相比，4种人结肠癌细胞系中miR-221表达均显著增高，差异具有统计学意义（P<0.01）；癌细胞转染anti-miR-221后，G0/G1期细胞比例明显增加，而S期细胞比例显著下降，并可见细胞凋亡的发生（P<0.01）。结论miR-221特异性抑制剂可通过抑制细胞增殖同时诱导凋亡而抑制结直肠癌细胞生长，提示miR-221作为结直肠癌生物治疗有效靶点有进
3、一步研究价值。关键词：结直肠癌；miRNA-221；增殖；凋亡中图分类号：R735.3文献标志码：A文章编号：1673-4254（2011）04-0674-04EffectofmiR-221-specificinhibitorontheproliferationandapoptosisofhumancolorectalcarcinomacellsWANGWei,SUNKai,WUCheng-tang,LEIShang-tong,ZENGJun-jie,WUYing-jun,LIGuo-xinDepartmentofGeneralSurgery,NanfangHospital,So
4、uthernMedicalUniversity,Guangzhou510515,ChinaAbstract:ObjectiveToinvestigatemiRNA-221expressioninhumancolorectalcarcinoma(CRC)cellsandtheeffectsofmiR-221-specificinhibitorontheproliferationandapoptosisofCRCcells.MethodsFourhumanCRCcelllines(HT-29,Lovo,SW-480,andCaCO2)wereexaminedformiRNA-221e
5、xpressionusingreal-timeQ-PCR.Thespecific2,-methoxy-modifiedRNAoligonucleotidesofmiR-221(anti-miR-221)weresynthesizedandtransfectedintoCaco2cellsvialiposome,andthechangesintheexpressionofmiR-221inthecellsweredetectedbyreal-timeQ-PCR.TheproliferationandapoptosisofthetransfectedCRCcellsweredetec
6、tedusingMTTassayandflowcytometry.ResultsThe4humanCRCcellsshowedsignificantlyupregulatedexpressionofmiR-221comparewithHUVECs(P<0.01).ThemiR-221-specificinhibitor,anti-miR-221,significantlyinhibitedtheexpressionofmiR-221inCaco2cellsandsuppressedthecellproliferation,causingalsoobviouscellapoptos
7、is(P<0.01).ConclusionThemiR-221-specificinhibitorshowspotentinhibitoryeffectonthegrowthofCRCcells,suggestingitsvalueasapotentialanti-tumorcandidatefortreatmentofCRC.Keywords:colorectalcarcinoma;miRNA-221;proliferation;apoptosis肿瘤的危害对人类生命和健康是不
