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时间:2018-08-23
《survivin 反义寡核苷酸诱导胃癌原代细胞凋亡的实验研究.doc》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、survivin反义寡核苷酸诱导胃癌原代细胞凋亡的实验研究【摘要】目的探讨survivin反义寡核苷酸对人胃癌原代细胞凋亡的影响,为胃癌的靶向治疗提供实验依据。方法利用人胃癌实体瘤新鲜标本培养胃癌原代细胞,胃癌细胞分为空白对照组、脂质体组、正义寡核苷酸组及反义寡核苷酸转染组。用脂质体介导survivin反义寡核苷酸转染胃癌原代细胞,48h后,观察肿瘤细胞的形态变化,Westernblot检测survivin蛋白的表达,流式细胞术检测细胞的凋亡。结果反义寡核苷酸转染组肿瘤细胞体积变小,细胞碎裂,出现凋亡小体;survivin蛋白量下降;流式细胞术检测到肿瘤细胞的凋亡率增高。与其他各组相比
2、,其差异有统计学意义(P<0.05),而空白对照组、脂质体组、正义寡核苷酸组之间的差异无统计学意义(P>0.05)。结论survivin反义寡核苷酸在脂质体介导下转染人胃癌原代细胞,可以诱导肿瘤细胞发生凋亡,抑制细胞生长,反义survivin可能成为一个促进胃癌细胞凋亡的手段。【关键词】survivin;反义寡核苷酸;凋亡;原代细胞;胃癌Researchoneffectsofsurvivinantisenseoligonucleotideinducingapoptosisofprimarycellofhumangastriccarcinoma[Abstract]Object
3、iveToinvestigatetheeffectsofsurvivinantisenseoligonucleotideoninducingapoptosisofprimarycellofgastriccarcinomaandprovidetheevidenceinexperimentforthetargetingtherapyofgastriccarcinoma.MethodsTheprimarycellswereculturedwiththefreshtissuefromhumangastriccarcinoma.Theculturedprimarycellswerecollect
4、edanddividedintofourgroups:shamcontrolgroup,liposomescontrolgroup,SODNcontrolgroupandASODNexperimentgroup.Aftertransfectingcellsfor48hoursusingsynthesizedsurvivinASODNbyliposome,culturedcellwereharvestedtocarryoutnexttests:primarycellmorphologicalchangeswereobservedbytheinvertedmicroscope,surviv
5、inproteinexpressionwasdetectedbyWesternblot,cellapoptosiswasexaminedbyflowcytometry.ResultsPrimarycellsweretransfectedafter48hours,abnormalmorphologicalchangeofprimarycellwasobservedinASODNtransfectedgroup.TheexpressionofsurvivinproteininASODNgroupwasdecreasedascomparedwithothercontrolgroupsbyWe
6、sternblotdetection.TheflowcytometryshowedtheARofASODNgroupwassignificantlyhigherthanthatofothergroups.TherewerestatisticaldifferencesbetweenASODNgroupandtheothergroups(P<0.05),andnodifferencesamongshamgroup,lipsomegroupandlip-SODNgroup(P>0.05).ConclusionSurvivinantisenseoligonucleotidecani
7、nducetheapoptosisandinhibitthegrowthoftheprimarycellofgastriccarcinoma.SurvivinASDONmightbeamethodto6promotetheapoptosisofthecellofgastriccarcinoma.[Keywords]survivin;antisenseoligonucleotide;apoptosis;primarycell;gastriccar
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