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时间:2020-05-04
《CIP1基因表达对BEL-7402肝癌细胞增殖和凋亡的影响-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在应用文档-天天文库。
1、·1534·2014年5月27日第94卷第2O期NatlMedJChina,May!.丝:.肝癌诊治.小RNA靶向激活p21WAn基因表达对BEL一7402肝癌细胞增殖和凋亡的影响吴志明黄欢刘卓炜郭睁垮蒋丽娟董培贾国金陈刚【摘要】目的探讨RNA激活技术上调p21基因的表达而影响人肝癌细胞株BEL-7402的增殖和凋亡能力。方法将与p21基因启动子区域DNA序列互补的RNA分子(dsP21—322)转染入BEL一7402细胞中,采用qPCR法及Western印迹方法检测p21“的表达;采用WST一1细胞增殖试剂检测细胞增殖情况;使用流式细胞仪评估肿瘤细胞凋亡情
2、况并使用Western印迹方法检测相关分子表达水平的变化。结果dsP21—322转染72h后,BEL一7402细胞中p21”表达显著上调;细胞增殖受到明显抑制,凋亡率36.86%(细胞的体积变小、变形,细胞核固缩,染色质凝聚呈深染,细胞出现皱缩、变圆、脱落)较对照组(11.51%、】4.06%)相比明显升高。结论RNA激活技术靶向上调p21’基因表达并抑制细胞增殖、增加诱导肿瘤细胞凋亡,可作为肝细胞癌或其他恶性肿瘤基因治疗的一种有效手段。【关键词】癌,肝细胞;细胞增殖;p21“Activationofp21bysmallactivatingRNAinhibi
3、tscellproliferationandinducesapoptosisinBEL-7402hepatomacellWuZhiming,HuangHuan,LiuZhuowei,GuoZhengzheng,JiangL~uan,DongPei,JiaGuojin,ChenDepartmentofUrology,CancerCenter,SunYat—senUniversity,StateKeyLaboratoryofOncologyinSouthernChina,Guangzhou,Guangzhou510060,ChinaCorrespondingau
4、thor:ChenGn,DepartmentofUrology,JinshanHospital,FudanUniversity,Shanghai201508,China,Email:chgan305@163.com【Abstract】ObjectiveToevaluatetheantineoplasticeffectsofp21transcriptionalactivationinducedbyduplexRNAsinhepatocellulm’carcinonla(HCC)celllineBEL一7402.MethodsCe11sweretreatedwi
5、thdsRNAscomplementarytopronl0tersequeneesofp21w“.Quantitativepolymerasechainreaction(qPCR)andWesternblotwereemployedtodetecttheexpressionofp21.Atvarioustimepointspost—transfection,cellviabilityassayandapoptosisanalysiswereusedtodeterminetheeffectofRNAactivation.AtiertransfectionWes
6、ternblotwasa]soperformedtodetecttheexpressionofBcl—xL,cleavedcaspase一3.cleavedcaspase-9andcleavedPARP.ResultsDsP21—322transfectionsignificantlyinhibitedcellviability.And.atDay5.dsP21—322inhibitedcellgrowthby65.84%versuscontro1.FlowcytometryrevealedthatdsP21—322causedasignificantinc
7、reaseofcellapoptosis.Thetotalpercentofapoptoticcells(UR+LR)increasedto36.86%versus11.51%and14.06%inmocksandcontrolSrespectively.Suchphenomenacorrelatedwithadecreaseofanti—apoptoticproteinBcl—xLandanincreaseofc】cavedcaspase一3.cleavedcaspase一9andcleavedPARP.ConclusionActivationofp21g
8、eneexpressionbysaRNAmayoff
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