姜黄素通过诱导Egr-1基因表达抑制HepG-2细胞的增殖  

Curcumin Inhibited HepG-2 Cell Proliferation through Inducing Egr-1 Gene Expression

作　　者：虞游[1] 蒋汉伟 卢佳伟[1] 陈建明[1] 

机构地区：[1]杭州师范大学生命与环境科学学院,杭州310036

出　　处：《中国细胞生物学学报》2015年第6期840-845,共6页Chinese Journal of Cell Biology

基　　金：<细胞生物学>市级精品课程(批准号:ZX13007002001);杭州师范大学实验室开放项目(批准号:SYSKF2014001);杭州师范大学"本科教学创一流工程"(批准号:PD11009009008)资助的课题~~

摘　　要：利用姜黄素处理肝癌Hep G-2细胞研究其对细胞增殖的影响以及可能的分子机制。姜黄素处理后,半定量RT-PCR分析Hep G-2细胞内肿瘤相关基因的表达水平,对筛选得到的姜黄素诱导表达上调的Egr-1基因作进一步验证。随后,设计Egr-1 sh RNA并构建p Green Puro-Egr-1重组质粒,通过转染细胞,构建稳定干扰细胞系。然后,在干扰细胞系的基础上分析Egr-1表达沉默在姜黄素对Hep G-2细胞增殖影响中的作用。结果发现,不同浓度姜黄素处理24 h或48 h后,Hep G-2细胞增殖明显下降并呈剂量效应关系,20μmol/L姜黄素处理不同时间后,其增殖也明显下降并呈时间效应关系;Egr-1基因在姜黄素处理后表达明显升高;Egr-1表达沉默显著减弱了姜黄素抑制Hep G-2细胞增殖的能力。上述结果提示,姜黄素抑制Hep G-2细胞增殖可能是通过诱导Egr-1基因表达介导的,表明其在肝癌的治疗中具有一定的应用前景。Hep G-2 cells were treated with curcumin to investigate its effect on cell proliferation and the underlying molecular mechanism. After curcumin treatment, expression level of tumor-related genes was analyzed. The up-regulated gene Egr-1 was concerned and its expression in curcumin-treated Hep G-2 cells was verified. Subsequently, Egr-1 sh RNA was designed and constructed into p Green Puro vector to obtain the recombinant plasmid p Green Puro-Egr-1. The stable interfering cell lines were established by puromycin selection. Finally, the resistant cell lines were used to analyze the role of Egr-1 interference in Hep G-2 cells exposed to curcumin. It was found that Hep G-2 cell proliferation was decreased in a dose-dependent manner when treated with different concentrations of curcumin for 24 or 48 h. When treated with 20 μmol/L of curcumin, the proliferation was decreased in time-dependent manner. Furthermore, Egr-1 gene expression was dramatically elevated when Hep G-2 cells were treated with curcumin. Interestingly, the suppression effect of curcumin in Hep G-2 cell proliferation was attenuated when Egr-1 gene expression was interfered. Based on all of the above results, it is concluded that curcumin inhibits Hep G-2 cell proliferation through inducing Egr-1 gene expression, and it might have the prospect in treating hepatocarcinoma.

关 键 词：姜黄素 HEPG-2细胞 增殖 EGR-1 RNA干扰 

分 类 号：R285[医药卫生—中药学]