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目的:探讨反义Survivin核酸(anti-pcDNA3-svv)对胃癌细胞系SGC7901的凋亡诱导、对泰索帝的化疗增敏作用及对化疗耐药的逆转作用.方法:采用脂质体转染法转染胃癌细胞系SGC7901,并筛选阳性克隆,以Westernblot检测Survivin蛋白的表达,RT-PCR检测MDR-1mRNA的表达,用透射电镜及TUNEL法观察并检测转染后SGC7901的凋亡变化,采用MTT法检测转染细胞对泰索帝的敏感性.结果:转染anti-pcDNA3-svv的SGC7901细胞系(SGC7901-SVVanti)其Survivin蛋白比未转染者明显降低,仅为其29.9%(P<0.01);透射电镜下可见转染组细胞呈凋亡晚期变化,TUNEL显示转染组与未转染组AI分别为0.241和0.083(P<0.01);转染组MDR-1mRNA较未转染组明显降低,转染组与未转染组MDR指数分别为:0.196±0.013和3.126±0.019(P<0.01);转染组对泰索帝的IC50值为16.7±1.98μg/L,而未转染组为55.7±1.89μg/L,差异具有显著的统计学意义(P<0.01).结论:反义Survivin核酸可诱导SGC7901细胞凋亡,增强泰索帝化疗敏感性,逆转耐药.
Objective: To investigate the apoptosis-inducing effect of anti-pcDNA3-svv on gastric cancer cell line SGC7901, the chemosensitization to Taxotere, and the reversal effect of chemosensitivity. Methods: Transfection with Lipofectamine The gastric cancer cell line SGC7901 was transfected and positive clones were screened. The expression of Survivin protein was detected by Western blot. The expression of MDR-1 mRNA was detected by RT-PCR. The apoptosis of SGC7901 after transfection was detected by transmission electron microscope and TUNEL method. The sensitivity of transfected cells to Taxotere was detected by MTT assay. Results: The Survivin protein of the SGC7901 cell line (SGC7901-SVVanti) transfected with anti-pcDNA3-svv was significantly lower than that of the untransfected cells (29.9%). <0.01); Under the transmission electron microscope, the cells in the transfection group showed late apoptosis, TUNEL showed that the AI of the transfected group and the untransfected group were 0.241 and 0.083 respectively (P<0.01); the MDR-1 mRNA in the transfection group was not transfected. The group’s MDR was significantly lower in the transfected group and the non-transfected group: 0.196±0.013 and 3.126±0.019, respectively (P<0.01); the IC50 value of the transfection group to Taxotere was 16.7±1.98 μg/L. The transfection group was 55.7±1.89μg/L, the difference was statistically significant (P<0.01). Conclusion: Antisense Survivin nucleic acid can induce SGC7901 Apoptosis enhances the sensitivity of Taxotere and reverses drug resistance.