Survivin反义寡核苷酸诱导胃癌细胞凋亡的研究

Effects of survivin antisense oligonucleotide on inducing apoptosis of human gastric cancer cell

目的探讨生存素反义寡核苷酸(ASODN)对人胃癌细胞SGC7901凋亡的诱导作用。方法设计合成特异性靶向survivin ASODN。胃癌细胞株SGC7901分为4组:空白对照组(Sham)、单纯脂质体对照组(Lip)、正义链转染对照组(Lip-SODN)、ASODN转染组(Lip-ASODN)。作用12、24、48 h后收获各组细胞。Western blot法检测各组细胞survivin表达情况,倒置显微镜观察细胞生长形态变化,流式细胞仪检测各组细胞凋亡率,免疫组化SP法检测细胞中Ki67表达。结果脂质体介导survivin ASODN转染后的胃癌细胞出现survivin蛋白表达明显下降;形态学上表现为细胞膜起泡、染色质固缩、凋亡小体形成;细胞凋亡率明显高于各对照组(P<0.05);细胞中Ki67表达水平明显降低。结论survivin反义寡核苷酸转染胃癌细胞能下调survivin蛋白表达,诱导胃癌细胞凋亡,抑制细胞增殖,具有明显的抗癌作用。

Objective This study was designed to investigate the apoptosis effects of survivin antisense oligonucleotide（ASODN） on human gastric carcinoma cell SC, C7901. Methods ASODN targeting surviving was designed and constructed. Cultured cells were divided into 4 groups： control group, lipofectin group, sense oligonucleotide（SODN） group, and ASODN group. After transfection for 12, 24, 48 h, cultured cells were harvested to carry on the next tests. Survivin protein expression was detected by Western blot; the apoptosis of SGC7901 cells was studied by the morphological observation using inverted microscope; apoptotie rate （AR） was examined by flow cytometry; the expression of Ki67 was detected in gastric cancer cells by immunohistochemical staining. Results The expression of survivin in SC, C7901 cells decreased after being transfected with survivin ASODN; Typical characteristics of apoptotic cells were found in the gastric cancer cell line, including bubbles on the membrane, condensed chromosome and apoptotic bodies; AR of ASODN group was significantly higher than that of other groups; the test by immunohistochemical method indicated that transfected ASODN downregulated the expression levels of Ki67. Conclusions Expression of survivin may decrease in gastric cancer cells after ASODN transfection. ASODN targeting survivin can induce cell apoptosis and inhibit cell proliferation. Survivin ASODN is a prospective anti-cancer drug.