冬凌草甲素抑制肝癌细胞HepG-2生长的实验研究

目的 探讨冬凌草甲素诱导肝癌细胞株HepG-2凋亡的作用及其机制.方法 MTT法检测细胞存活率;Annexin V和PI染色后流式细胞术检测HepG-2细胞凋亡率;Hoechst染色后荧光显微镜观察细胞形态;Western blotting检测MAPK信号通路凋亡相关蛋白的表达.结果 冬凌草甲素作用HepG-2细胞24 h后,可剂量依赖性减少细胞存活率,半数抑制率IC50为38.41 μmol/L.高、中、低浓度冬凌草甲素处理24 h后的流式结果显示,随浓度增加细胞凋亡率明显增加,HepG-2细胞可见典型的凋亡核改变.Western blotting结果证实冬凌草甲素可降低HepG-2细胞Bcl-2、caspase-9和caspase-3的蛋白表达,增加p-JNK、p-p38、p-p53和活化的caspase-9的蛋白表达.结论 冬凌草甲素可诱导肝癌HepG-2细胞凋亡,MAPK信号转导通路凋亡相关蛋白的活化与冬凌草甲素诱导HepG-2细胞凋亡相关.

Inhibition of oridonin in HepG-2 cells proliferation

Objective To investigate the proapoptotic effect and tile underlying mechanism of oridonin in human hepatocarcinoma （HepG-2） cells. Methods MTr assay was used to measure the cell viability. Apoptosis was determined by flow cytometry analysis after Anncxin V/PI staining and fluorescence microscope after Hocchst staining.The expression of MAPK signaling pathway proteins was determined by immunoblot analysis. Results Treatment with oridonin for 24 h resulted in a marked decrease in cell viability of HepG-2 cells in a dose- dependent manner.The ICs0 values were determined as 38.41 p, mol/L for 24 h.Afler treatment with 20,40 and 60 p, mol/L oridonin for 24 h, apoptotic rates of HepG-2 ce]ls increased by flow cytometry analysis and typical apoptotic nucleus alterations were observed with fluorescence microscope.Treatment with 20,40 and 60 Ixmol/L oridonin down-regulated protein expression of Bcl-2, caspase-9, caspase-3 and increased the expression of p- JNK, p-p38,p-p53 as well as activated the cleavage of caspase-9.Conclusion Oridonin may induce the apoptosis of HepG-2 cells through MAPK pathway.