NS-398对人肝癌BEL-7402细胞凋亡作用及其机制的研究

NS-398 Regulation on Apoptosis in Hepatocullar Carcinoma BEL-7402 Cell Lines

目的:本研究应用NS-398对肝癌细胞株进行药物干预,探讨NS-398对肝细胞癌凋亡的影响。方法:NS-398取0、25、50、75、100μmol/L5个浓度组,每个浓度组选取24h、48h、72h三个作用时间点。MTT法测定生长抑制情况,FCM检测细胞周期、凋亡率和Survivin、cyclinD1、PTEN的表达。结果:NS-398可以显著抑制BEL-7402细胞增殖,使G0/G1期比例显著降低;G2/M期比例显著增高并且诱导细胞凋亡(P<0.001),并呈时间和剂量依赖性。与对照组相比,NS-398使Survivin、cyclinD1、PTEN蛋白表达显著下调并呈时间和剂量依赖性关系(P<0.001)。结论:NS-398可诱导BEL-7402细胞凋亡,可能部分通过下调Survivin和PTEN途径实现的。

Objective： To apply NS-398 for medicinal intervention of the cell lines of liver cancer and to investigate the influence of NS-398 on apoptosis of hepatocellular carcinoma. Methods： The media containing various concentrations of NS-398 （0, 25, 50, 75 and 100μmol/L） were used for concentration-dependent experiments. Three time-points （24h, 48h, 72h） of action time were taken for each concentration group, and the cells were collected daily for 3 days. The inhibition effect of cell growth was measured using MTT assay. Changes of cell eyc, le, the apoptotic cells and expression of survivin, cyclin D1 and PTEN after treatment with NS-398 were detected by flow cytometry method. Results： Treatment of with NS-398 significantly suppressed cell proliferation in time- and dose-dependent manner both. The percentage of cells in G0/G1 phase decreased as the percentage of cells in G2/M phase significantly increased in cultures treated with NS-398. NS-398 also induced the apoptosis detected by flow cytometry method. We found that NS-398 treatment significantly downregulated the expression of survivin, cyclin D1 and PTEN in a dose-dependence and time-dependent manner （P〈0.001）. Conclusion： This study suggests that NS-398 can inhibited the proliferation and induced apoptosis of hepatoma cell, which may be partially fulfilled by downregulation of the survivin and PTEN.