NS-398对肝癌SMMC7721和BEL-7402细胞凋亡和survivin、PTEN表达调节作用的研究

Effects of NS-398 on apoptosis of hepatic carcinoma SMMC-7721 and BEL-7402 cells and expression of survivin and PTEN

目的:研究应用NS398对肝细胞癌细胞株进行药物干预,探讨NS398对肝细胞癌细胞增殖和凋亡的影响。方法:人肝细胞癌细胞株SMMC7721、BEL7402细胞常规培养。NS398取0、25、50、75、100μmol/L5个浓度组,每个浓度组选取24、48、72h三个作用时间点进行检测,观察NS398对各细胞株的作用。MTT法测定2种细胞株的增殖抑制情况,FCM检测对细胞周期和凋亡的影响,FCM定量检测COX2、survivin、PTEN的表达。结果:MTT法结果显示,NS398可以显著抑制SMMC7721、BEL7402细胞增殖。FCM检测结果显示,NS398分别使SMMC7721、BEL7402细胞G0/G1期比例显著降低;G2/M期细胞比例显著增高,S期细胞比例无明显变化,并引起了显著的细胞凋亡(P<0.001),并且呈时间依赖性和剂量依赖性。与对照组相比,NS398使COX2、survivin和PTEN蛋白表达显著下调并且呈时间和剂量依赖性关系(P<0.001)。结论:NS398通过诱导SMMC7721、BEL7402细胞株凋亡而抑制其增殖,可能部分通过下调survivin和PTEN途径实现的。

Objective： NS-398 is a specific inhibitor of cyclooxygenase-2 （COX-2）, which is associated with proliferation, differentiation,apoptosis and carcinogenesis. In the present study, we aimed to analyze the inhibitory effects of NS-398 on the proliferation and apoptosis of hepatoma cells in vitro. Methods： Hepatic carcinoma SMMC-7721 and BEL-7402 cells were cultured. They were treated with NS-398 0,25,50,75,and 100 μmol/L, respectively. Each concentration group was subdivided into three groups： treatment for 24, 48, and 72 h, respectively. The cells were harvested daily for 3 days. Growth inhibition of the two cell lines were measured by MTT assay. Cell cycle,apoptosis, and expression of COX-2, survivin, and PTEN （phosphatase and tension homology deleted from chromosome 10） were determined quantitatively by flow cytometry （FCM） analysis. Results： Compared with control group,NS-398 significantly inhibited the proliferation of SMMC-7721 and BEL-7402 cells. NS-398 also significantly decreased the proportion of cells in the G0/G1 phase and increased the proportion of cells in the G2/M phase but had so significant effects on the proportion of cells in S phase. The expressions of COX-2, survivin, and PTEN were significantly down-regulated in SMMC-7721 and BEL-7402 cells by NS-398. All the effects of NS-398 were in a dose- and concentration-dependent manner（P〈0. 001）. Conclusion..NS-398 inhibited the proliferation of SMMC-7721 and BEL-7402 cells by inducing apoptosis of the two cells. The effects may be due to the down-regulation of expression of survivin and PTEN.