乌索酸通过抑制miR-21表达诱导肝癌HepG2细胞凋亡的机制
Mechanism study on apoptosis of hepatocellular carcinoma HepG2 cells induced by ursolic acid through inhibiting the expression of miR-21
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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。Abstract: This study aimed at investigating the effect of miR-21 on the apoptosis of hepatocellular carcinoma HepG2 cells induced by ursolic acid(UA). MTT 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 MTT 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.
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Keywords:
- ursolic acid /
- hepatocellular carcinoma cells /
- miR-21 /
- proliferation /
- apoptosis
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