摘要
探讨乌索酸通过调控miR-21表达从而诱导肝癌HepG2细胞凋亡的作用机制。采用MTT方法检测乌索酸对肝癌细胞增殖的抑制作用;qPCR检测肝癌细胞中miR-21的表达水平及乌索酸对HepG2细胞中miR-21表达的调控作用;转染miR-21 mimics进HepG2细胞中上调miR-21的表达后,MTT、流式细胞检测法、RT-PCR方法分别分析miR-21在乌索酸对细胞的增殖、凋亡以及对凋亡相关基因的调控过程中的作用。结果显示,与肝正常细胞L-02以及肝癌SMCC-7721、Bel-7402细胞相比较,乌索酸对肝癌HepG2细胞的增殖抑制效果最强,且HepG2细胞中miR-21的表达水平最高。乌索酸可下调HepG2细胞中miR-21的表达,且在24 h的下调作用最强。miR-21的表达上调可以部分抵消乌索酸对HepG2细胞的抑制增殖及促进凋亡,部分抵消下调凋亡抑制基因Bcl-2、survivin表达和上调促凋亡基因Bax表达的作用。结果提示,乌索酸通过抑制miR-21的表达诱导肝癌HepG2细胞凋亡。
This study aimed at investigating the effect of miR-21 on the apoptosis of hepatocellular carcinoma HepG2 ceils induced by ursolic acid （ UA）. MTF assay was used to determine the inhibition effect of ursolic acid on proliferation of hepatocellular carcinoma cells. The expression level of miR-21 in hepatocellular carcinoma cells and the regulation effect of ursolic acid on the expression of miR-21 in HepG2 cells were determined by qPCR. To up-regulate the expression of miR-21, miR-21 mimics were transfected into HepG2 cells. Then MTr assay, flowcytometry（ Annexin V-FITC staining）, and RT-PCR were used to detect the regulation effects of ursolic acid on the proliferation, apoptosis, and the expression of apoptosis-related genes after miR-21 over-expression. The results showed that the proliferation inhibition effect of ursolic acid on HepG2 cells and the expression level of miR-21 in HepG2 cells were higher than in hepatic cell L-02 and hepatocellular carcinoma SMCC-7721, Bel- 7402 cells. So further study was performed in the HepG2 cells. Ursolic acid inhibited the expression of miR-21 in HepG2 cells. And the greatest inhibition effect was at 24 h after treatment with UA. Over-expression of miR-21 partially offset the effects of ursolic acid on the proliferation, apoptosis, and the expression of apoptosis-related genes such as Bcl-2, survivin and Bax after 24 h. The results suggested that apoptosis of hepatocellular carcinoma HepG2 cells could be induced by ursolic acid by down-regulating the expression of miR-21.
出处
《中国药科大学学报》
CAS
CSCD
北大核心
2015年第6期745-750,共6页
Journal of China Pharmaceutical University
基金
宁夏自然科学基金资助项目(No.NZ14077)~~
关键词
乌索酸
肝癌细胞
MIR-21
增殖
凋亡
ursolic acid
hepatocellular carcinoma cells
miR-21
proliferation
apoptosis
