苦参碱对人肝癌细胞HepG2凋亡和PEG10表达的影响

Effects of matrine on the apoptosis and the expression of PEG10 in human hepatocarcinoma cell Line HepG2

目的:观察苦参碱(MAT)对人肝癌HepG2细胞凋亡及对PEG10表达的影响。方法:用MTT法检测不同质量浓度的MAT作用HepG2细胞后对细胞增殖的影响;JC-1染色流式细胞术检测对细胞线粒体膜电位的影响;RT-PCR和免疫细胞化学法分别检测对PEG10基因和蛋白水平的表达变化。结果:MAT质量浓度≥0.1 g/L时有抑制HepG2细胞增殖的作用,且作用随药物质量浓度的增加及时间的延长而加强(P<0.01);JC-1染色流式细胞术检测分析显示MAT作用前后HepG2细胞线粒体膜电位下降明显(P<0.01);RT-PCR及免疫细胞化学结果显示,经MAT处理后,PEG10的基因和蛋白表达水平均下调。结论:苦参碱能有效抑制PEG10的表达,且能抑制HepG2细胞增殖并可通过改变线粒体膜电位诱导细胞凋亡。

Objective To study the effects of matrine（MAT） on the apoptosis and the expression of PEG10 in human hepatocarcinoma cell line HepG2. Methods MTT assay was used to determine the proliferationinhibition activity by MAT to HepG2 cell. JC-1 staining was prepared to detect the change of mitochondrial membrane potential in HepG2 cells after MAT was given. RT-PCR and immunocytochemical method for detecting the PEG10 gene and protein expression levels were used. Results MAT could inhibit the HepG2 cell proliferation above the concentration of 0.125 mg / mL（different from above--MAT ≥ 0.1 g / L） and in a concentrationdependent and time-dependent manner（P〈0.01）. JC-1 staining and flow cytometry detection showed that MAT can significantly decrease the mitochondrial membrane potential of HepG2 cells（P 〈 0.01）. The RT-PCR and immunocytochemical staining results showed that 0.5 and 2.0 mg / ml（different from Chinese） MAT could reduce PEG10 gene and protein expression obviously. Conclusion MAT could decrease the expression level of PEG10 gene and inhibited cell proliferation,change the mitochondrial membrane potential and induce HepG2 cell apoptosis.