NK-104对人肝癌细胞凋亡及半胱氨酸蛋白酶3活性的影响

目的:研究日本新近研制的第三代3-羟基-3-甲基戊二酰辅酶A(3-hydroxy-3-methyl-glutaryl-CoA,HMG-CoA)还原酶抑制剂匹伐他汀(pitavastatin,NK-104)对人肝癌细胞凋亡及半胱氨酸蛋白酶3(caspase-3)活性的影响。方法:采用细胞培养技术,以肝癌细胞系HepG2为靶细胞,以不同浓度的药物处理细胞48 h后,利用WST-8法测定NK-104对细胞增殖的影响;利用荧光染料Hoechst 33258染色,荧光显微镜观察细胞核碎片;以流式细胞仪分析细胞周期的变化;采用半胱氨酸蛋白酶3比色法检测caspase-3活性。结果:NK-104(10μmol/L)对HepG2细胞有明显抑制作用,可诱导HepG2细胞凋亡,并能增强caspase-3基因的活性。结论:NK-104能够诱导HepG2细胞凋亡,其机制与caspase-3依赖性凋亡调节信号通路有关。

Effects of NK-104 on apoptosis and caspase-3 activity in hepatocellular carcinoma cells

OBJECTIVE: To study the apoptosis of human hepatocellular carcinoma cell line HepG 2 induced by pitavastatin (NK-104) and to investigate the change of caspase-3 activity. METHODS: HepG 2 cells were treated with NK-104 for 48 hours and observed under a microscope by using Hoechst 33258 staining. The proliferation of the cells was detected with WST-8 method. The apoptosis peaks were examined by flow cytometry. The caspase-3 activity was detected with caspase-3 colorimetric protease assay. RESULTS: The treatment of HepG 2 with 10 micromol/L NK-104 could inhibit the proliferation, and induce HepG 2 apoptosis and up-regulate the caspase-3 activity. CONCLUSION: NK-104 at 10 micromol/L induces apoptosis of HepG 2 cells. It is dependent on caspase-3 pathway in human hepatocellular carcinoma cells.