冬凌草甲素通过激活ATR通路诱导HEPG_2细胞凋亡实验研究

Oridonin Induced the HEPG_2 Cells Apoptosis by the Activation of the ATR Pathway

目的:探讨冬凌草甲素抑制HEGP_2肝癌细胞增殖及其机制研究方法:MTT法检测抑制率、结晶紫检测冬凌草甲素诱导HEPG_2肝癌细胞抑制增殖作用、Western blotting检测不同浓度冬凌草甲素作用HEPG_2肝癌细胞后ATR、H_2AX、γ-H_2AX、P53等蛋白的变化。结果:MTT法、结晶紫法显示冬凌草甲素对HEPG_2肝癌细胞能够明显抑制,且抑制作用在一定浓度范围内呈计量依赖性;Western blotting结果显示冬凌草甲素在诱导细胞凋亡过程中ATR、P-P53、γ-H_2AX表达水平及活性显著增强。结论:冬凌草甲素诱导HEPG_2肝癌细胞发生凋亡可能是通过引起H_2AX蛋白磷酸化进一步激活ATR信号通路引起细胞凋亡。

Objective： To research oridonin induce the HEPG2 cells apoptosis and its the related mechanism. Methods ： Using MTF and crystal violet staining to study oridonin inhibited HEPG2 cells proliferative, Western Blotting to detect the change of ATR, H2AX, γ-H2AX, P53 and so on protein after oridonin treated HEPG2 cells. Results： MTT method and crystal violet staining show that oridonin can significantly inhibited HEPG2 cells, and the inhibition effect in certain concentrations with a dose- dependent manner. Western blotting showed that the ATR, P-P53, γ-H/AX protein activity and express has significant enhanced during oridonin induced the HEPG2 cells apoptosis. Conclusion ： Oridonin induced the HEPG2 cells apoptosis may be cause by the phosphorylation of H2AX protein further activate the ATR signaling pathways lead to cell death.