重楼皂苷Ⅰ对肝癌细胞的增殖及凋亡的影响

Growth and Apoptosis Effects of Paris Saponin Ⅰ on Human Hepatocellular Carcinoma Cells

探讨重楼皂苷Ⅰ(paris saponinⅠ,PSⅠ)在体外对人肝癌SMMC-7721细胞株增殖和凋亡的影响及相关机制.MTT法检测PSⅠ对肝癌SMM-C7721细胞株的增殖抑制作用,Hoechst 33528染色法观察细胞核的形态学变化,流式细胞术Propidium iodide(PI)染色检测细胞周期及凋亡的情况,免疫印迹的方法检测Fas、Bcl-2、Bax、细胞周期素D1(cell cycle regulatory factor D1,Cyclin D1)和细胞周期素E(cell cycle regulatory factor E,Cy-clin E)的表达情况.PSⅠ能时间和浓度依赖性的抑制肝癌SMMC-7721细胞的增殖,与对照组比较,差异有统计学意义(P<0.05).干预后的SMMC-7721细胞染色质浓缩,核碎裂,凋亡小体形成,呈典型的凋亡变化.细胞周期阻滞于G1期,并且有凋亡峰形成,呈浓度依赖性.PSⅠ能浓度依赖性地上调Fas和Bax的表达,下调Bcl-2、Cyclin D1和Cyclin E蛋白的表达水平.PSⅠ可能是通过阻滞肿瘤细胞的生长及诱导细胞凋亡等机制,从而抑制肝癌细胞的增殖.

To investigate the effect of Paris Saponin Ⅰ（PSⅠ） on the growth and apoptosis of hepatoma car-cinoma cell strain（SMMC-7721） and its mechanism in vitro.The proliferation-inhibiting effect of PSⅠ on SMMC-7721 cells was measured by MTT assay while the nuclear morphology of apoptotic cells was assessed by Hoechst 33528 staining.Cell cycle and apoptosis were examined by flow cytometry.Protein expressions of Fas,Bax,Bcl-2,cell cycle regulatory factor D1（Cyclin D1） and cell cycle regulatory factor E（Cyclin E） were detected by Western blot analysis.The results showed that PSⅠ inhibited the growth of SMMC-7721 cells in a dose and time-dependent manner in vitro.Marked morphological changes,including conden-sation of chromatin,nuclear fragmentation and apoptotic bodies were clearly shown on Hoechst 33528 staining.Flow cytometry analysis showed PSⅠ arrested SMMC-7721 cells in G0/G1 phase and induced apoptosis.PSⅠ treatment increased the Fas and Bax protein expressions while decreased the protein expressions of Bcl-2,Cyclin D1 and cyclin E.PSⅠ can inhibit the proliferation of hepatoma cells by prolonging G1 period of cell cycle,and inducing cell apoptosis.