survivin反义寡核苷酸对肝癌细胞生长抑制作用的研究  

Inhibitory effect of antisense oligonucleotide on the expression of survivin gene and proliferation of human hepatocellular carcinoma cell line SMMC-7721

作　　者：陈涛[1] 贾玉容[1] 赵铁军[1] 尹致良[1] 

机构地区：[1]中国人民解放军成都军区总医院全军普通外科中心,四川省成都市610083

出　　处：《世界华人消化杂志》2004年第7期1546-1549,共4页World Chinese Journal of Digestology

摘　　要：目的:采用反义寡核苷酸封闭肝癌细胞中survivin基因的表达,研究其对肝癌细胞生长的抑制作用. 方法:针对survivin mRNA翻译起始位点设计并合成反义寡核苷酸(antisense oligonucleotide,ASODN);采用脂质体介导survivin ASODN转染人肝癌细胞株SMMC-7721细胞,western-blot及原位杂交方法检测survivin蛋白及mRNA表达,流式细胞仪检测细胞凋亡率的变化,检测转染前后细胞贴壁率及细胞生长抑制率变化,绘制细胞生长曲线. 结果:脂质体介导survivin反义寡核苷酸转染肝癌细胞后survivin蛋白及mRNA表达分别从69.59及75.61降低至10.71及22.94,细胞贴壁率从90.68%降低至33.16%, 细胞凋亡率从0.7%增加至31.35%,均有显著差异.转染后对细胞生长的抑制作用可以维持大约1 wk,在转染后第3 d抑制率最高可达71.8%. 结论:脂质体介导转染survivin反义寡核苷酸可以有效降低细胞内survivin基因的表达,诱导细胞发生凋亡,抑制肝癌细胞生长.AIM: To investigate the inhibitory effect of survivin antisense oligonucleotide (ASODN) on the expression of survivin gene and proliferation of human hepatocellular carcinoma cell line SMMC-7721. METHODS: The 20mer antisense oligonucleotide (ASODN) targeted to the promotor region of survivin mRNA was designed and synthesized. The expression of survivin gene in hepatocellular carcinoma cell line SMMC-7721 was blocked by means of ASODN transfection mediated by DOTAP liposomal reagent. The changes of survivin protein and mRNA expression after transfection were assessd by Western blot and in situ hybridization, respectively. The apoptotic rate was detected by flow cytometer. The changes of cell adherent rate, cell growth activity, and the inhibitory rate of cell growth were also studied. RESULTS: The expression of survivin protein and mRNA was decreased markedly after survivin ASODN transfection (69.59 vs 10.71, and 75.61 vs 22.94, P <0.01). Meanwhile, the cell adherent rate also decreased markedly (90.68% vs 33.16%, P<0.01) while the apoptotic rate increased markedly (0.7% vs 31.35%, P<0.01). The inhibitory effect on the cell growth could maintain about a week after transfection, and the highest inhibitory rate could be 71.8% three days after transfection. CONCLUSION: Transfection of ASODN targeted to the pro-motor region of survivin mRNA by DOTAP liposomal transfection reagent can down-regulate the expression of survivin protein and mRNA significantly in 7721 cell line and inhibit the proliferation of cancer cells, survivin may be an important target in the therapy of hepatocellular carcinoma.

关 键 词：SURVIVIN 反义寡核苷酸 肝癌 癌细胞 生长抑制作用 

分 类 号：R735.7[医药卫生—肿瘤]