miR-101 reverses hypomethylation of the PRDM16 promoter to disrupt mitochondrial function in astrocytoma cells.
miR-101 reverses hypomethylation of the PRDM16 promoter to disrupt mitochondrial function in astrocytoma cells.
复制标题
miR-101 逆转 PRDM16 启动子的低甲基化，从而破坏星形细胞瘤细胞中的线粒体功能。
DOI：
10.18632/oncotarget.6652
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发表时间：
2016-01-26
期刊：
影响因子：
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通讯作者：
Wu M
中科院分区：
文献类型：
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作者：
Lei Q;Liu X;Fu H;Sun Y;Wang L;Xu G;Wang W;Yu Z;Liu C;Li P;Feng J;Li G;Wu M
我们在先前的研究中发现了PR结构域基因家族中的一个成员PRDM 16，它是星形细胞瘤中一个新的甲基化相关基因。这项先前的研究也报道了miR-101是胶质瘤中的肿瘤抑制因子。本研究证实，PRDM 16是一个低甲基化基因，可以在星形细胞瘤患者中过表达，并表明PRDM 16启动子的低甲基化状态可以预测星形细胞瘤患者的预后不良。本文报道的结果表明，PRDM 16直接被miR-101抑制，也通过表观遗传调控被抑制。PRDM 16被证实是miR-101的新靶点，并被miR-101直接抑制。miR-101还通过改变PRDM 16启动子的甲基化状态来降低PRDM 16的表达。miR-101与PRDM 16启动子上甲基化相关组蛋白H3 K4 me 2和H3 K27 me 3的减少以及H3 K9 me 3和H4 K20 me 3的增加相关。此外，EZH 2、EED和DNMT 3A被鉴定为miR-101的直接靶标，并且miR-101通过靶向DNMT 3A抑制PRDM 16表达，DNMT 3A降低PRDM 16核心启动子处的组蛋白H3 K27 me 3和H3 K4 me 2。本文报道的结果表明，miR-101破坏细胞线粒体功能并通过线粒体途径诱导细胞凋亡;例如，MMP和ATP水平降低，而ADP/ATP比率和ROS水平、裂解的Caspase-9和裂解的PARP水平、Bax/Bcl-2比率和Smac从线粒体释放到细胞质的水平增加。PRDM 16的敲低逆转了miR-101抑制的抗凋亡作用。总之，miR-101逆转了PRDM 16启动子的低甲基化，这抑制了PRDM 16的表达，破坏了细胞线粒体功能，并诱导了细胞凋亡。
Our previous report identified PR domain containing 16 (PRDM16), a member of the PR-domain gene family, as a new methylation associated gene in astrocytoma cells. This previous study also reported that miR-101 is a tumor suppressor in glioma. The present study confirms that PRDM16 is a hypomethylated gene that can be overexpressed in astrocytoma patients and demonstrates that the hypomethylation status of the PRDM16 promoter can predict poor prognoses for astrocytoma patients. The results reported herein show that PRDM16 was inhibited by miR-101 directly and also through epigenetic regulation. PRDM16 was confirmed as a new target of miR-101 and shown to be directly inhibited by miR-101. miR-101 also decreased the expression of PRDM16 by altering the methylation status of the PRDM16 promoter. miR-101 was associated with a decrease in the methylation-related histones H3K4me2 and H3K27me3 and an increase in H3K9me3 and H4K20me3 on the PRDM16 promoter. In addition, EZH2, EED and DNMT3A were identified as direct targets of miR-101, and miR-101 suppressed PRDM16 expression by targeting DNMT3A which decreases histone H3K27me3 and H3K4me2 at the PRDM16 core promoter. The results reported here demonstrate that miR-101 disrupted cellular mitochondrial function and induced cellular apoptosis via the mitochondrial pathway; for example, MMP and ATP levels decreased, while there was an increase in ADP/ATP ratios and ROS levels, levels of cleaved Caspase-9 and cleaved-PARP, the Bax/Bcl-2 ratios, and Smac release from the mitochondria to the cytoplasm. Knockdown of PRDM16 reversed the anti-apoptotic effect of miR-101 inhibition. In summary, miR-101 reversed the hypomethylation of the PRDM16 promoter which suppressed the expression of PRDM16, disrupted cellular mitochondrial function, and induced cellular apoptosis.