苦参碱诱导人肝癌细胞系HepG_2凋亡的研究

Study on apoptosis of human hepatoma cell line HepG_2 induced by matrine

目的 观察苦参碱是否能诱导人肝癌细胞系HepG2 凋亡及凋亡相关基因的变化。方法 0～ 3 .0g/L苦参碱处理HepG2 细胞 ,光镜和电镜下观察细胞形态 ;TUNEL法原位检测DNA断裂 ;流式细胞仪检测凋亡峰 ;AnnexinV FITC/PI双标记检测早期凋亡 ;免疫组化和原位杂交法检测wp5 3 ,bcl 2 ,bax ,c myc ,Fas等凋亡相关基因的表达 ,采用真彩色图像分析仪定量检测。结果 1.5～ 3 .0g/L苦参碱可诱导HepG2 凋亡 ,并且使凋亡相关基因wp5 3 ,bax ,Fas表达上调 ,而抗凋亡基因bcl 2 ,c myc表达下调。结论 一定浓度的苦参碱可诱导人肝癌细胞HepG2 凋亡 ,并且该凋亡与凋亡相关基因表达增强和抗凋亡相关基因表达减弱有关。

Objective To observe the apoptosis of human hepatoma cell line HepG 2 induced by Matrine and to investigate the changes of apoptosis associated genes. Methods HepG 2 cells were treated with 0 3.0 g/L matrine for 3 days, and observed under optical microscope and electron microscope. DNA break was detected with TUNEL in situ assay, the apoptosis peaks were detected with flow cytometry, the early stage apoptosis was detected with Annexin V FITC/PI double labelled assay, the expression of apoptosis associated genes wp53, bcl 2, c myc, bax and Fas were detected with immunohistochemical technique and in situ hybridization , all results were quantitatively analyzed with high fidelity colored picture dissector.Results 1.5 3.0 g/L Matrine could induce HepG 2 apoptosis and up regulate the expression of apoptotic genes wp53, bax, Fas and down regulate the expression of anti apoptotic genes bcl 2, c myc. Conclusion Matrine in certain concentrations can induce apoptosis of HepG 2 cell and the apoptosis is related to the up regulation apoptotic genes and down regulation of anti apoptotic genes.