花椒提取物诱导肝癌细胞凋亡作用和分子机制研究

摘要

目的：研究花椒提取物ZBME诱导的肝癌细胞凋亡作用及其分子机制。方法：体外培养肝癌细胞HepG2细胞株。分为对照组（生理盐水）和ZBME处理组。不同浓度的ZBME处理体外培养肝癌细胞，通过相差倒置显微镜观察对照组和处理组细胞失巢凋亡和生长抑制。免疫印迹（ Western blotting）方法检测ZBME对细胞内抗凋亡蛋白Mcl-1、Survivin和Bcl-xL以及促凋亡蛋白Bax表达调控，基于Annexin V-PI双染的流式细胞分析统计 ZBME 诱导的细胞凋亡率。结果：在相差显微镜下，可以观察到 ZBME 能够显著诱导HepG2肝癌细胞株发生细胞凋亡和生长抑制。MTT检测细胞相对活性结果显示ZBME能诱导HepG2细胞增殖抑制。8μg/ml ZBME处理细胞48h增殖抑制率约40%（P﹤0.05）。Annexin V-PI染色表明ZBME能诱导细胞显著凋亡，8μg/ml ZBME 处理细胞48h凋亡诱导率约90%（P﹤0.05）。结论：ZBME能够诱导肝癌细胞凋亡和增殖抑制。ZBME抗癌作用可能通过下调致癌蛋白Mcl-1、Survivin和Bcl-xL以及上调抑癌蛋白Bax实现。%Objective:To investigate the effects of Zanthoxylum bungeanum Maxim extract( ZBME)-induced ap-optosis in hepatocellular carcinoma cells. Methods:HepG2 cells were cultured in vitro. We set up control group and ZBME-treated group. Cells untreated or treated with various conditions were observed under contrast microscope to show apoptosis and cell growth inhibition. MTT assay was performed to measure cell viability. Western blotting was performed to detect apoptosis-related molecules such as Mcl-1,Survivin,Bcl-xL,Bax. FACs based on Annexin V-PI double staining was used to assess apoptosis ratio. Results:ZBME induced apparent apoptosis and cell growth in-hibition in HepG2 cells. Apoptosis and cell growth inhibition induced by ZBME may be mediated by down-regulation of anti-apoptotic proteins Mcl-1,Survivin and Bcl-xL,and up-regulation of pro-apoptotic protein Bax. Conclu-sion:ZBME effectively exerted apoptotic cytotoxicity in liver cancer cells by regulating apoptotic protein expression.