MiR-138 Induces Renal Carcinoma Cell Senescence by Targeting EZH2 and Is Downregulated in Human Clear Cell Renal Cell Carcinoma
MiR-138 Induces Renal Carcinoma Cell Senescence by Targeting EZH2 and Is Downregulated in Human Clear Cell Renal Cell Carcinoma
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DOI：
10.3727/096504013x13775486749218
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发表时间：
2013-01-01
影响因子：
3.1
通讯作者：
Zhao, Jun
中科院分区：
文献类型：
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作者：
Liang, Jiaqian;Zhang, Yajing;Zhao, Jun
MiR-138已被证明在各种癌症中下调，包括头颈部鳞状细胞癌（HNSCC）和透明细胞肾癌（ccRCC）。本研究旨在揭示miR-138诱导肾癌细胞衰老的机制并鉴定其特异性靶基因。我们使用qRT-PCR分析肾癌细胞系和ccRCC样品中的miR-138表达水平。在miR-138模拟物转染后测量β-半乳糖苷酶的活性用于功能分析。为了确定miR-138的靶点，我们使用三种类型的靶点预测软件确定了三个候选靶基因。此外，进行3 'UTR荧光素酶测定。Western blotting检测候选靶基因的蛋白表达水平。此外，进行EZH 2的siRNA敲低。与对照组相比，RCC细胞系和肿瘤样品中miR-138的表达下调。转染miR-138模拟物可诱导SN-12细胞衰老，降低EZH 2蛋白表达，增加P16蛋白表达。此外，miR-138降低了EZH 2的3 'UTR荧光素酶活性。siRNA敲低EZH 2基因可诱导SN-12细胞衰老，EZH 2蛋白表达水平降低，P16蛋白表达水平升高。miR-138是ccRCC中的肿瘤抑制miRNA，其通过下调EZH 2表达和上调P16表达来诱导SN-12细胞衰老。
MiR-138 has been shown to be downregulated in various cancers, including head and neck squamous cell carcinoma (HNSCC) and clear cell renal carcinoma (ccRCC). In the present study, we aimed to reveal the mechanism of miR-138 induction of senescence in renal carcinoma cells and identify its specific target genes. We used qRT-PCR to analyze miR-138 expression levels in renal carcinoma cell lines and ccRCC samples. The activity of beta-galactosidase was measured for functional analysis after miR-138 mimic transfection. To identify the targets of miR-138, we used three types of target prediction software to determine three candidate target genes. Furthermore, a 3'UTR luciferase assay was performed. Western blotting was used to detect the protein expression levels of candidate target genes. Additionally, knockdown of EZH2 by its siRNA was performed. The expression of miR-138 was downregulated in RCC cells lines and in tumor samples compared with their controls. Transfection of miR-138 mimic induced SN-12 cell senescence, decreased the protein expression of EZH2, and increased the protein expression of P16. Furthermore, miR-138 decreased the 3'UTR luciferase activity of EZH2. The knockdown of EZH2 by siRNA induced SN-12 cell senescence, decreased the protein expression level of EZH2, and increased the protein expression of P16. MiR-138 is a tumor-suppressor miRNA in ccRCC that induces SN-12 cell senescence by downregulating EZH2 expression and upregulating P16 expression.