siRNA腺病毒载体沉默MEKK3基因促进TRAIL诱导肝癌HCC-9204细胞凋亡-论文.pdf

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1、0，o一．Z0．【论著】siRNA腺病毒载体沉默MEKK3基因促进TRAIL诱导肝癌HCC．9204细胞凋亡薛同春，葛宁灵，孙瑞霞，赵燕(复旦大学附属中山医院肝癌研究所，复旦大学癌变与侵袭原理教育部重点实验室，上海200032)摘要：目的研究抑制丝裂原活化蛋白激酶／细胞外信号调节激酶一激酶3(MEKK3)基因表达促进TRAIL诱导人肝癌HCC一9204细胞凋亡的作用。方法构建人MEKK3基因的siRNA重组腺病毒载体，转导人肝癌HCC一9204细胞，以RT-囫PCR和印迹法检测MEKK3mRNA和蛋白的表达，

2、筛选并建赢稳定沉默MEKK3基因表达的人肝癌HCC一9204细胞株，以流式细胞仪检测TRAIL诱导细胞凋亡的情况。结果①成功构建并鉴定表达人MEKK3基因的siRNA重组腺病毒载体。②建赢MEKK3基因有效且稳定沉默的人肝癌HCC一9204细胞株。③500g／LTRAIL处理MEKK3基因沉默的HCC一9204细胞株，细胞凋亡率较野生HCC·9204细胞株显著提高。结论本研究成功建立有效且稳定沉默MEKK3基因表达的人肝癌HCC一9204细胞株模型，初步证实MEKK3基因表达与TRAIL诱导肝癌细胞凋亡的敏感

3、性相关。关键词：肝癌细胞系；丝裂原活化蛋白激酶／细胞外信号调节激酶一激酶3(MEKK3)；TRAIL；siRNA；腺病毒载体中图分类号：R735．7文献标志码：A文章编号：1672—9188(2013)12—0720—07MEKK3siRNArecombinantadenovirusvectortransductioninducesapoptosisofhumanlivercancercelllineHCC一9204withTRAILXUETong-chun，GENing—ling，SUNRui—xia，ZH

4、AOYan(LiverCancerInstitute,ZhongshanHospitalFudanUniversity,Shanghai200032,"MinistryofEducationKeyLaboratoryforCarcinogenesisandCancerInvasion，FudanUniversity,Shanghai200032，China)Abstract：ObjectiveToinvestigatethepromotingefectofsilencingMEKK3geneonTRAIL．i

5、nducedapoptosisoflivercancerHCC·9204cells．SOastosearchforanoveltreatmentforlivercancer．MethodsMEKK3siRNArecombinantadenovirusvectorwasconstructedandtransducedinto1ivercancerHCC．9204cells．RT-PCRandWesternblottingwasperformedtoanalyzethesuppressionofMEKK3inth

6、etransducedcells．FlowcytometrywasusedtomeasureTRAIL．inducedapoptosis．Resul~DNAsequencingandRT-PCRconfirmedthattheMEKK3siRNAexpressionadenovirusvectorsweresuccessfullyconstructed．RT-PCRandWesternblottingshowedthatMEKK3stablyandefectivelysilencedHCC一9204cells

7、wereestablished．ThesignificantincreasingofTRAIL(500gg／L)-inducedapoptosiswasobservedinMEKK3silencedHCC一9204cellsbyflowcytometryanalysis．ConclusionTheMEKK3stablyandefectivelysilencedlivercancerHCC一9204cellsweresuccessfullyestablishedthecorrelationbetweenMEKK

8、3expressionandTRAIL—inducedapoptosiswasdemonstrated．TheMEKK3silencedHCC一9204cellsprovideatoolforfurtherinvestigationofthemechanism．KeyWords：livercancercellline：mitogen·activatedproteinkinase／extraeellu