miR-495 promotes apoptosis and inhibits proliferation in endometrial cells via targeting PIK3R1
miR-495 promotes apoptosis and inhibits proliferation in endometrial cells via targeting PIK3R1
复制标题
miR-495通过靶向PIK3R1促进子宫内膜细胞凋亡并抑制增殖
DOI：
10.1016/j.prp.2019.01.020
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发表时间：
2019-03-01
影响因子：
2.8
通讯作者：
Ruan, Peng
中科院分区：
文献类型：
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作者：
Tan, Aili;Luo, Ruoyu;Ruan, Peng
子宫内膜癌（EC）是妇女健康的巨大威胁。本研究旨在探讨microRNA (miR)-495在体外培养EC细胞增殖和凋亡中的作用。采用逆转录-定量聚合酶链反应（RT-qPCR）检测mRNA水平。此外，双荧光素酶报告基因检测证实PIK3R1是miR-495的靶标。转染miR-495模拟物后，进行细胞计数试剂盒8 （CCK-8）检测，评估EC细胞的细胞活力。western blotting检测转染后PIK3R1、血管内皮生长因子（VEGF）、Bcl-2、Bax、caspase 3蛋白的表达。流式细胞术检测EC细胞凋亡率。这些结果表明，miR-495在肿瘤组织中较邻近正常组织明显下调，而PIK3R1上调。转染miR-495模拟物后，EC细胞的增殖明显受到抑制，凋亡明显促进。此外，转染miR-495 mimic后，观察到PIK3R1、Bcl-2、VEGF表达下调，Bax和caspase 3表达上调。综上所述，这些研究结果表明，miR-495在体外EC细胞的转录后水平上直接靶向PIK3R1，从而起到肿瘤抑制基因的作用。
Endometrial cancer (EC) is a huge threat to women's health. The aims of this study were to investigate the role of microRNA (miR)-495 in the proliferation and apoptosis of EC cells in vitro. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to detect the mRNA levels. In addition, dual-luciferase reporter assay was used to verified that PIK3R1 was a target of miR-495. After transfection with miR-495 mimics, Cell Counting Kit 8 (CCK-8) assay was performed to evaluate the cell viability of EC cells. The protein expression of PIK3R1, vascular endothelial growth factor (VEGF), Bcl-2, Bax, caspase 3 after transfection was analyzed using western blotting. Furthermore, cell apoptosis rate of EC cells was evaluated by flow cytometry. These results showed that miR-495 was significantly down-regulated in tumor tissues compared with the adjacent normal tissues, while PIK3R1 was up-regulated. The proliferation of the EC cells that were transfected with miR-495 mimics was markedly inhibited, and apoptosis was significantly promoted. In addition, down-regulated expression of PIK3R1, Bcl-2, VEGF expression and upregulated expression of Bax and caspase 3 expression were observed after transfected with miR-495 mimic. Together these findings indicated that miR-495 acts as a tumor suppressor gene by directly targeting PIK3R1 at the post-transcriptional level in EC cells in vitro.