凋亡抑制因子反义核酸对肝癌细胞生物学作用研究

目的探讨survivin反义寡核苷酸对人肝癌细胞凋亡、增殖的影响。方法设计合成特异性sur-vivin的反义寡核苷酸(ASODN)。以高表达survivin基因的人肝癌细胞系(SMMC-7721)为靶细胞、反义sur vivin(AS-ODN)为阻断剂,倒置显微镜观察细胞形态变化,WesternBlot法检测细胞survivin表达情况,MTT试验观察AS-ODN对该细胞系的生长抑制作用,流式细胞仪分析细胞增殖周期。结果AS-ODN明显抑制了SMMC-7721肝癌细胞的生长,细胞分裂阻滞在分裂期的中期,并诱导细胞发生凋亡,而各对照组细胞生长良好,细胞增殖指数明显低于各对照组(P<0.05)。结论survivingASODN能下调其蛋白表达,诱导人肝癌细胞凋亡,抑制细胞增殖。

Biological effect of survivin antisense olignucleotide in human hepatocarcinoma cells

Objective To explore biological effect of survivin antisense olignucleotide on the cell growth and apoptosis of human hepatocarcinoma. Methods ASODN targeting survivin gene was designed and sythesised. Using survivin gene over expression hepatocarcinoma cell line SMMC-7721 as target cell and survivin AS-ODN as gene expression blocking agent, cell growth and mitotic character was analyzed by MTT experiment and Flow cytometry, cell morphological change was observed under microscope and survivin expression was detected by western blot. Results Expression of survivin in ASODN treated hepatocarcinoma cells was significantly blocked compared with that in control group and the growth of hepatocarcinoma cells were suppressed by AS-ODN. The SMMC-7721 cell cycle was arrested in metaphase and cell apoptosis was found and abnormal morphological changes was observed in these cells. Conclusion Survivin ASODN can not only effectively block survivin gene expression in hepatocarcinoma cells but also induce these cells apoptosis, which means that survivin gene may become a new target for the hepatocarcinoma bio-treatment.