雷帕霉素对人肝癌细胞BEL-7402体外抗肿瘤作用研究

The Studies of Antitumor Effect of Rapamycin on Human Hepatocellular Carcinoma BEL-7402 Cells in vitro

目的:观察雷帕霉素体外对肝癌细胞BEL-7402生长、细胞凋亡以及细胞转移能力作用。方法:以5nmol/L,10nmol/L,20nmol/L,30nmol/L,40nmol/L和50nmol/L不同浓度的雷帕霉素作用于体外培养的BEL-7402细胞,MTT法检测细胞生长抑制率,应用流式细胞仪检测细胞凋亡,Hoechst33258荧光染色法观察细胞凋亡时的形态学变化,细胞粘附分析及体外侵袭试验观察肿瘤细胞的转移能力。结果:雷帕霉素可显著的抑制BEL-7402细胞的生长,诱导细胞发生凋亡,并呈现出明显的量-效与时-效关系。雷帕霉素作用肝癌细胞BEL-740248h后,在Hoechst33258荧光染色图片上可见核浓缩及核碎裂等典型的细胞凋亡特征。雷帕霉素能抑制其粘附性和侵袭性,具有剂量依赖性,随浓度增加而抑制增加。结论:雷帕霉素不仅能诱导BEL-7402肝癌细胞凋亡,抑制细胞的生长,而且同时抑制肿瘤细胞的粘附性和侵袭性,阻止肿瘤细胞的转移,雷帕霉素可能成为一种潜在的抗肝癌药物。

Objective： To investigate the effect of cell growth, cell invasion and apoptosis of rapamycin on human hepatocelluar carcinoma BEL-7402 cells. Methods： Different concentrations of rapamycin, such as 5nmol/L, 10nmol/L, 20nmol/L, 30nmol/L, 40nmol/L and 50nmol/L, were given to the in vitro cultured BEL-7402 cells. Cell viability was assessed by MTF assay, cell apoptosis was observed using Hoechst 33258 staining and flow cytometry （FCM）. The cell adhesion and invasive effect were investigated by the standard methods. Results： Rapamycin could inhibit the growth of BEL-7402 cells and can significantly cause apoptosis, both suppression and apoptosis occurred in a time-and dose-dependent manner. Marked morphological change of cell apoptosis was observed by Hoechst 33258 staining; rapamycin inhibited the adhesion and the invasiveness in a concentration-dependent manner. Conclusion： Rapamycin has a significant anti-proliferation effect by induction of apoptosis, and can prevent metastasis by inhibition of the adhesion and invasiveness.