EZH2 promotes hepatocellular carcinoma progression through modulating miR-22/galectin-9 axis.
EZH2 promotes hepatocellular carcinoma progression through modulating miR-22/galectin-9 axis.
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EZH2通过调节miR-22/galectin-9轴促进肝细胞癌进展
DOI：
10.1186/s13046-017-0670-6
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发表时间：
2018-01-09
期刊：
影响因子：
--
通讯作者：
Li H
中科院分区：
文献类型：
--
作者：
Chen S;Pu J;Bai J;Yin Y;Wu K;Wang J;Shuai X;Gao J;Tao K;Wang G;Li H
近期研究表明，干扰素-γ（IFN-γ）诱导库普弗细胞中半乳凝素-9的表达，在调节肝炎相关肝细胞癌（HCC）的微环境中起着至关重要的作用。然而，IFN-γ是否会诱导肝癌细胞中半乳凝素-9的表达，以及其在肝癌发生发展中的生物学作用和调控机制尚不明确。
采用定量聚合酶链反应（qPCR）和蛋白质免疫印迹分析检测经IFN-γ刺激的肝癌细胞系中半乳凝素-9和EZH2的水平。利用生物信息学分析和荧光素酶报告基因实验，证实miR - 22与半乳凝素-9的3′非翻译区（3’-UTR）结合。通过甲基化特异性PCR（MSP）和亚硫酸氢盐测序PCR（BSP）分析miR - 22启动子的甲基化状态，同时采用染色质免疫沉淀（ChIP）实验确定EZH2和H3K27me3在启动子上的占据状态。此外，分别使用CCK - 8法、Transwell实验和流式细胞术分析，评估半乳凝素-9和miR - 22异位表达对细胞增殖、迁移、侵袭和细胞凋亡的影响。
IFN - γ诱导肝癌细胞系中半乳凝素-9和EZH2的上调。半乳凝素-9是miR - 22的靶标，EZH2通过使miR - 22启动子上的H3K27发生三甲基化，而非使DNA处于高甲基化状态，来促进半乳凝素-9的表达。miR - 22过表达在体外和体内均抑制肝癌细胞的生长、侵袭和转移。有趣的是，半乳凝素-9也表现出抗肿瘤作用，在miR - 22过表达的细胞中恢复半乳凝素-9的表达，可增强其抗肿瘤作用。
这些研究结果表明，EZH2通过表观遗传抑制miR - 22来促进半乳凝素-9的表达，并且作为免疫抑制剂的半乳凝素-9在肝癌中也发挥着肿瘤抑制作用。
本文的网络版本（10.1186/s13046 - 017 - 0670 - 6）包含补充材料，仅授权用户可获取。
Recent studies have shown that interferon-γ (IFN-γ)-induced galectin-9 expression in Kupffer cells plays an essential role in modulatingthe microenvironment of hepatitis-associated hepatocellular carcinoma (HCC). However, whether or not IFN-γ induces galectin-9 expression in HCC cells, its biological role and regulatory mechanism in HCC development and progression are poorly defined. Quantitative PCR and western blotting analysis were used to detect galectin-9 and EZH2 levels in HCC cell lines stimulated with IFN-γ. Bioinformatics analysis and luciferase reporter assay were utilized to confirmthe binding ofmiR-22 to the 3′ untranslated region (3’-UTR) of galectin-9. The methylation status of miR-22 promoter was analyzed by MSP (Methylation specific PCR) and BSP (bisulfite sequencing PCR), while chromatin immunoprecipitation (ChIP) assay identify the occupation status of EZH2 and H3K27me3 at the promoter. Furthermore, the effect of ectopic expression of galectin-9 and miR-22 on cell proliferation, migration, invasion and cell apoptosis was assessed by using CCK-8, transwell assays and flow cytometric analysis, respectively. IFN-γ induces up-regulation of galectin-9 and EZH2 in HCC cell lines. Galectin-9 is a target of miR-22 and EZH2 facilitates galectin-9 expression by tri-methylation of H3K27 on miR-22 promoter but not hyper-methylation status of DNA. MiR-22 overexpression suppressed HCC cell growth, invasion, and metastasis both in vitro and in vivo. Interestingly, galectin-9 also exhibited antitumor effects, and restoring galectin-9 expression in miR-22 overexpressing cells strengthened its antitumor effects. These findings indicated that EZH2 facilitates galectin-9 expression by epigenetically repressing miR-22 and that galectin-9, which is known as an immunosuppressant, also functions as a tumor suppressor in HCC. The online version of this article (10.1186/s13046-017-0670-6) contains supplementary material, which is available to authorized users.
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50.3
作者：
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