Ki-67反义寡核苷酸抑制人肝癌细胞株HEPG-7402生长的体外试验

Antisense Oligodeoxyribonucleotide Target Ki-67 Inhibits Proliferation of Human Hepatocellucar Carcinoma Cell HEPG-7402 in Vitro

目的通过观察硫代磷酸化修饰的Ki-67抗原反义寡核苷酸(antisenseoligodeoxyribonucleoti-de,ASODN)抑制Ki-67抗原表达,从而抑制HEPG-7402细胞体外增殖,为今后肝癌的基因治疗提供新的手段。方法将Ki-67反义寡核苷酸(ASODN)作用于HEPG-7402细胞,MTT比色法测细胞增殖活性;免疫细胞化学(ImmunocellulerchemistryICC)方法检测Ki-67标记指数(LabelingindexLI)、RT-PCR方法观察Ki-67mRNA水平的改变。结果Ki-67ASODN作用组HEPG-7402细胞增殖受到明显抑制;Ki-67标记指数(LI)下降,Ki-67mRNA合成减少。结论针对Ki-67的反义寡核苷酸能抑制人肝癌细胞株的体外生长。

Objective To investigate phosphorothioated antisense oligodeoxynucleotide （ASODN） target Ki-67 decreasing the antigene＇s expression and inhibiting proliferation of HEPG-7402 cell in vitro. It will show us a new method to cure hepatocellular carcinoma by genetherapy in the future. Methods We treat HEPG-7402 cells with ASODN and observe changes of cell＇s proliferation by the way of MTT. ICC technique was used to find the change of cell＇s Ki-67 labeling index （LI）. RT-PCR method was used for detecting Ki-67 mRNA＇s change. Results HEPG-7402 cell＇s growth was inhibited, Ki-67 labeling index was decreased, and the expression of Ki-67 mRNA was decreased in ASODN group. Conclusion ASODN target against Ki-67 can inhibit the proliferation of human hepatocellular carcinoma cell HEPG-7402 in vitro.