蓝萼甲素对人肝癌细胞HepG2细胞株的作用

The effects of glaucocalyxin A in hepatic cancer HepG2 cell line

目的观察蓝萼甲素对人肝癌细胞HepG2的抑制和诱导细胞凋亡作用。方法不同浓度蓝萼甲素处理HepG2细胞后,MTT检测24h,48h细胞生长抑制率,LDH试剂盒检测细胞LDH释放量,倒置显微镜观察细胞形态,RT-PCR法对凋亡相关基因Bc-l 2,bax和c-myc的mRNA表达进行检测,流式细胞仪检测细胞凋亡率。结果在1.25~20μmol.L-1剂量范围内,蓝萼甲素对HepG2的生长有显著抑制作用,并呈剂量依赖性。蓝萼甲素5,10,20μmol.L-1能够使HepG2培养液中的LDH释放量显著增加,倒置相差显微镜观察,可见细胞明显皱缩。同时蓝萼甲素可使细胞Bc-l2表达减少,Bax表达增加,c-myc表达减少。结论蓝萼甲素可抑制HepG2细胞增殖并诱导其凋亡,其机制可能与调控Bc-l2,Bax,c-myc的mRNA的表达有关。

OBJECTIVE To investigate the effect of of hepatocellular carcinoma HepG2 cell line. METHODS glaucocalyxin A （GLA） on cell proliferation and apoptosis HepG2 cells were treated with different concentrations of GLA.The cell proliferation inhibition of HepG2 cells by GLA was observed by MTT assay after 24 and 48 hours. Intracellular LDH release were determined by LDH assay. The morphosis of HepG2 was observed by inverted phase contrast microscope. The expression of Bcl-2, Bax and C-myc were detected by RT-PCR.The cell apoptosis of HepG2 cells exposed to GLA was analyzed by flow cytometry. RESULTS Within 1.25-20μmol·L^-1, GLA obviously inhibited the proliferation of HepG2 cells in a dose-dependent manner. 5, 10, 20μmol· L^-1 GLA could significantly increase intracellular LDH releasing to culture medium（P〈 0.01）. Cytoplasm vacuolization could be observed by inverted phase contrast microscope. GLA could increase the expression of Bax mRNA, and decrease the expression of Bcl-2 and C-myc mRNA. In the whole, GLA could induce apoptosis. CONCLUSION GLA can obviously inhibit HepG2 cell proliferation and induce cell apoptosis. The mechanism of this apoptotic effect is closely related to regulation of expression of Bcl-2, Bax, C-mye mRNA.