survivin 反义寡核苷酸诱导胃癌原代细胞凋亡的实验研究.doc

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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