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时间:2020-05-20
《转染 PDCD5质粒促进顺铂诱导 A549细胞凋亡作用的研究-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、重庆医学2014年9月第43卷第26期3449论著·基础研究doi:10.3969/j.issn.1671—8348.2014.26.015转染PDCD5质粒促进顺铂诱导A549细胞凋亡作用的研究戴玲丽,梅同华,张明川。(1.南昌大学第一附属医院呼吸内科,南昌330006;2.重庆市铜梁县人民医院呼吸内科402560)摘要:目的观察程序化死亡因子5(PDCD5)基因联合顺铂诱导人肺腺癌A549细胞凋亡的作用,探索其可能机制。方法利用脂质体瞬时转染PDCD5重组质粒入A549细胞,RT-PCR检测其转染效率。Westernblot检测转染后PDCD5、Bcl-2和Survivin蛋白表达情况。
2、MTT法及流式细胞仪检测PDCD5单药和联合顺铂后的细胞凋亡情况。结果PDCD5重组质粒成功转染入A549细胞,Westernbolt结果显示转染重组质粒组中PDCD5蛋白表达高于空白对照组及转染空质粒组,Bcl一2和Survivin蛋白则低于空白对照组及转染空质粒组,差异有统计学意义(P<0.05)。MTT及流式细胞仪检测显示转染重组质粒组较空白对照组及转染空质粒组细胞凋亡率高(P3、4)26-3449-03TransfectionofPDCD5plasmidforpromotingcisplatin-inducedA549cellsapoptosisDaiLingli,MeiTqnghua1ix,ZhangMingchuan。(1.DepartmentofRespiration,FirstA,liatedHospital,SchoolofMedicine,NanchangUniversity,Nanchang,Jiangxi330006,China;2.DepartmentofRespiration,TongliangCountyPeoplesHospital,Chon4、gqing402560,China)Abstract:ObjectiveToobservetheroleofprogrammedcelldeath5(PDCD5)genecombinedwithcisplatinforinducingtheap—optosisofhumanlungadenocarcinomaA549cellsandtoinvestigateitspossiblemechanism.MethodsThePDCD5recombinantplas—midwastransientlytransfeetedintoA549cellsbylipofectamine.Itstransfe5、ctionefficiencywasdetectedbyRT-PCR.TheexpressionsoftrasfectedPDCD5protein.Bcl-2andSurvivinproteinwereexaminedbyWesternbolt.TheapoptosisofA549cellsbysinglePDCD5anditscombinationwithcisplatinwasmeasuredbytheMTTmethodandtheflowcytometry.ResultsPDCD5recombinantplasmidwastransfectintoA549cellssuccessful6、ly.TheWesternblotresultsshowedthattheexpressionofPDCD5proteininthetransfectionrecombinantplasmidgroupwashigherthanthatintheblankcontrolgroupandthetransfectionemptyplasmidgroup,whiletheexpres—sionofBcl_2andSurvivinproteinwaslowerthanthatintheothertwogroups,thedifferencewasstatisticallysignificant(P<7、0.05).TheMTTandflowcytometryresultsdemonstratedthatthecellapoptosisrateinthetransfectionrecombinantplasmidgroupwashigherthanthatintheblankcontrolgroupandthetransfectionemptyplasmidgroup(P<0.05).ConclusionPD
3、4)26-3449-03TransfectionofPDCD5plasmidforpromotingcisplatin-inducedA549cellsapoptosisDaiLingli,MeiTqnghua1ix,ZhangMingchuan。(1.DepartmentofRespiration,FirstA,liatedHospital,SchoolofMedicine,NanchangUniversity,Nanchang,Jiangxi330006,China;2.DepartmentofRespiration,TongliangCountyPeoplesHospital,Chon
4、gqing402560,China)Abstract:ObjectiveToobservetheroleofprogrammedcelldeath5(PDCD5)genecombinedwithcisplatinforinducingtheap—optosisofhumanlungadenocarcinomaA549cellsandtoinvestigateitspossiblemechanism.MethodsThePDCD5recombinantplas—midwastransientlytransfeetedintoA549cellsbylipofectamine.Itstransfe
5、ctionefficiencywasdetectedbyRT-PCR.TheexpressionsoftrasfectedPDCD5protein.Bcl-2andSurvivinproteinwereexaminedbyWesternbolt.TheapoptosisofA549cellsbysinglePDCD5anditscombinationwithcisplatinwasmeasuredbytheMTTmethodandtheflowcytometry.ResultsPDCD5recombinantplasmidwastransfectintoA549cellssuccessful
6、ly.TheWesternblotresultsshowedthattheexpressionofPDCD5proteininthetransfectionrecombinantplasmidgroupwashigherthanthatintheblankcontrolgroupandthetransfectionemptyplasmidgroup,whiletheexpres—sionofBcl_2andSurvivinproteinwaslowerthanthatintheothertwogroups,thedifferencewasstatisticallysignificant(P<
7、0.05).TheMTTandflowcytometryresultsdemonstratedthatthecellapoptosisrateinthetransfectionrecombinantplasmidgroupwashigherthanthatintheblankcontrolgroupandthetransfectionemptyplasmidgroup(P<0.05).ConclusionPD
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