survivin 反义寡核苷酸诱导胃癌原代细胞凋亡实验探究.doc

survivin 反义寡核苷酸诱导胃癌原代细胞凋亡实验探究 　【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，48 h后，观察肿瘤细胞的形态变化，Western blot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，其差异有统计学意义（Plt;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（Pgt;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 Research on effects of survivin antisense oligonucleotide inducing apoptosis of primary cell of human gastric carcinoma ［Abstract］ Objective To investigate the effects of survivin antisense oligonucleotide on inducing apoptosis of primary cell of gastric carcinoma and provide the evidence in experiment for the targeting therapy of gastric carcinoma.Methods The primary cells were cultured with the fresh tissue from human gastric carcinoma.The cultured primary cells were collected and divided into four groups:sham control group，liposomes control group，SODN control group and ASODN experiment group.After transfecting cells for 48 hours using synthesized survivin ASODN by liposome，cultured cell were harvested to carry out next tests:primary cell morphological changes were observed by the inverted microscope，survivin protein expression was detected by Western blot，cell apoptosis was examined by flow cytometry.Results Primary cells were transfected after 48 hours，abnormal morphological change of primary cell was observed in ASODN transfected group.The expression of survivin protein in ASODN group was decreased as compared with other control groups by Western blot detection.The flow cytometry showed the AR of ASODN group was significantly higher than that of other groups.There were statistical differences between ASODN group and the other groups （Plt;0.05），and no differences among sham group，lipsome group and lip-SODN group（Pgt;0.05）.Conclusion Survivin antisense oligonucleotide can induce the apo