黄连素对肝癌Hep-G2细胞株的作用研究

Effect and Mechanism of Berberine on Human Hepatocellular Carcinoma Cell Line Hep-G2

摘要: 目的：探讨黄连素(berberine, BBR)作用于肝癌Hep-G2细胞增殖和凋亡的机制。方法：CCK-8法检测BBR对Hep-G2细胞增殖的抑制作用；流式细胞术检测BBR对细胞凋亡的影响；蛋白印迹法(Western blotting)及实时荧光定量PCR (RT-PCR)检测抗增殖基因BTG2及细胞周期蛋白CyclinB1、CyclinD1的蛋白及mRNA表达的情况。结果：与对照组相比，BBR可以抑制肝癌Hep-G2细胞的增殖，且抑制率与BBR浓度成正比，差异具有统计学意义(p < 0.01)；与对照组相比，BBR可诱导细胞凋亡(p < 0.01)，且凋亡率随着黄连素的作用时间的延长而增加(p < 0.01)；BTG2 mRNA的表达随着BBR作用时间的延长逐渐增强(p < 0.01)，且与对照组相比，差异具有显著性(p < 0.01)，而CyclinB1、CyclinD1mRNA的表达则随着BBR作用时间的延长逐渐减弱(p < 0.01或p < 0.05)，与对照组相比，差异具有显著性(P < 0.01)；与对照组相比，BTG2和CyclinB1、CyclinD1蛋白表达的差异具有显著性(p < 0.05)。结论：BBR可抑制肝癌细胞Hep-G2的增殖，诱导细胞凋亡，其机制可能与其上调抗增殖基因BTG2和下调细胞周期蛋白CyclinB1、CyclinD1的表达有关。

Abstract: Objective: To investigate the effect and mechanism of Berberine on inhibiting cell proliferation and inducing apoptosis in Hepatocellular carcinoma Hep-G2 cell line. Methods: The inhibitory ef-fect of Berberine on the proliferation of Hep-G2 cells was detected by CCK-8 method; effect of Berberine on cell apoptosis by flow cytometry (FCM); western blotting and real time fluorescent quantitative PCR (RT-PCR) analysis were used to detect the mRNA and protein expression of anti proliferation gene BTG2 and CyclinB1, cyclin D1. Results: The proliferation of Hep-G2 cell lines was inhibited by Berberine in a dose and time dependent manner, and compared with the control group, the difference was statistically significant (p < 0.01). Berberine promotes cell apoptosis, and the rate of apoptosis is proportional to the effect time of Berberine (P < 0.01). The expression of BTG2 mRNA increased with the time of Berberine treatment (p < 0.01), and compared with the control group, there’s significant difference (p < 0.01); while the expression of CyclinB1, CyclinD1 mRNA was gradually decreased with the time of Berberine intervention (p < 0.01 or p < 0.05), and the difference was significantly compared with the control group (p < 0.01). Compared with the control group, the differential expression of BTG2 and CyclinB1, CyclinD1 protein was significant (p < 0.05). Conclusions: Berberine can inhibit the proliferation of Hep-G2 cells and induce cells apoptosis, which mechanism may be related to the up-regulation of BTG2 and down-regulation of CyclinB1, Cyclin D1 by berberine.