miR-6743-5p, as a direct upstream regulator of GRIM-19, enhances proliferation and suppresses apoptosis in glioma cells
miR-6743-5p, as a direct upstream regulator of GRIM-19, enhances proliferation and suppresses apoptosis in glioma cells
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miR-6743-5p作为GRIM-19的直接上游调节因子，增强神经胶质瘤细胞的增殖并抑制细胞凋亡
DOI：
10.1042/bsr20171038
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发表时间：
2017-12-22
影响因子：
4
通讯作者：
Yao, Shengtao
中科院分区：
文献类型：
--
作者：
Cao, Fang;Zhang, Qiang;Yao, Shengtao
维甲酸干扰素诱导死亡相关基因19（GRIM - 19）已被确认为一种肿瘤抑制蛋白，它通过信号转导和转录激活因子3（STAT3）信号通路以及神经胶质瘤细胞中的非STAT3通路调节细胞生长、凋亡和迁移。在此，我们研究了神经胶质瘤细胞中调节GRIM - 19表达的分子机制。通过TargetScan算法，确定了四种微小RNA（miRNA），即hsa - miR - 17 - 3p、hsa - miR - 423 - 5p、hsa - miR - 3184 - 5p和hsa - miR - 6743 - 5p，它们有可能与GRIM - 19的3' - 非翻译区（3' - UTR）结合。进一步的miRNA抑制剂转染和荧光素酶实验表明，miR - 6743 - 5p能够直接靶向GRIM - 19的3' - UTR。此外，在神经胶质瘤标本和细胞系中，miR - 6743 - 5p的表达与GRIM - 19的表达呈负相关。而且，抑制miR - 6743 - 5p可显著抑制神经胶质瘤细胞的增殖，并明显促进其凋亡，而这种表型可通过敲低GRIM - 19来挽救。最后，抑制miR - 6743 - 5p的表达可抑制STAT3的磷酸化，以及STAT3的两个靶基因细胞周期蛋白D1（CyclinD1）和B细胞淋巴瘤/白血病 - 2（Bcl - 2）的mRNA表达，而miR - 6743 - 5p模拟物则具有相反的作用。用STAT3抑制剂AG490处理可部分挽救miR - 6743 - 5p过表达或GRIM - 19敲低所产生的促进增殖和抗凋亡效应。总之，miR - 6743 - 5p可能通过靶向GRIM - 19和STAT3在神经胶质瘤中充当癌基因微小RNA（oncomiRNA）。
Gene associated with retinoid-interferon-induced mortality-19 (GRIM-19) has been recognized as a tumor suppressor protein, which regulates cell growth, apoptosis, and migration by signal transducer and activator of transcription 3 (STAT3) signaling pathway and non-STAT3 pathway in glioma cells. Here, we investigated the molecular mechanisms that regulated GRIM-19 expression in glioma cells. By the TargetScan algorithm, four miRNAs, hsa-miR-17-3p, hsa-miR-423-5p, hsa-miR-3184-5p, and hsa-miR-6743-5p, were identified with the potential to bind with 3′-UTR of GRIM-19. Further miRNA inhibitor transfection and luciferase assays revealed that miR-6743-5p was able to directly target the 3′-UTR of GRIM-19. Additionally, miR-6743-5p expression was inversely related with GRIM-19 expression in glioma specimens and cell lines. Moreover, the inhibition of miR-6743-5p caused a significant inhibition of cell proliferation and a marked promotion of cell apoptosis in glioma cells, and this phenotype was rescued by GRIM-19 knockdown. Finally, the inhibition of miR-6743-5p expression suppressed the phosphorylation of STAT3, and the mRNA expression of CyclinD1 and Bcl-2, two target genes of STAT3, while miR-6743-5p mimic had the inversed effects. Treatment with STAT3 inhibitor AG490 partially rescued the proliferation-promoting and anti-apoptosis effects of miR-6743-5p overexpression or GRIM-19 knockdown. Collectively, miR-6743-5p may act as an oncomiRNA in glioma by targetting GRIM-19 and STAT3.