miR-138 suppresses the proliferation, metastasis and autophagy of non-small cell lung cancer by targeting Sirt1.
miR-138 suppresses the proliferation, metastasis and autophagy of non-small cell lung cancer by targeting Sirt1.
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DOI：
10.3892/or.2017.5619
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发表时间：
2017-06
期刊：
影响因子：
4.2
通讯作者：
Cao Z
中科院分区：
文献类型：
--
作者：
Ye Z;Fang B;Pan J;Zhang N;Huang J;Xie C;Lou T;Cao Z
本研究探讨miR-138在非小细胞肺癌(NSCLC)中的作用及其机制。总共收集了45例新鲜切除的临床非小细胞肺癌组织。用实时荧光定量聚合酶链式反应检测miR-138在组织和细胞中的表达。体外将miR-138模拟物导入A549和CALU-3细胞，分别用CCK-8比色法、Transwell和流式细胞仪检测miR-138对肺癌细胞增殖、细胞周期和侵袭转移的影响。免疫印迹法检测潜在靶基因Sirt1在肺癌细胞中的蛋白表达。用双荧光素酶报告基因分析进一步确认Sirt1是否是miR-138的靶基因。MiR-138在肺癌组织中的表达明显降低，且与肺癌的分化程度和有无淋巴结转移呈负相关。体外实验结果表明，miR-138能抑制肺癌细胞的增殖、侵袭和迁移。已证实miR-138可下调Sirt1蛋白表达，抑制上皮-间充质转化(EMT)，降低AMPK信号通路活性，上调mTOR磷酸化水平。双荧光素酶报告实验证明miR-138可以直接调控Sirt1。单独下调Sirt1也可以引起同样的分子和生物功能变化。Western印迹分析和共聚焦显微镜结果表明，miR-138过表达或干扰Sirt1表达可能通过AMPK-mTOR信号通路抑制肺癌细胞自噬活性。MIR-138在肺癌中发挥肿瘤抑制作用。它可能通过下调Sirt1的表达和激活细胞自噬来抑制肺癌的增殖、侵袭和转移。MiR-138基因表达下调与肺癌的发生发展密切相关。
The present study determined the role and mechanism of miR-138 in non-small cell lung cancer (NSCLC). In total, 45 freshly resected clinical NSCLC tissues were collected. The expression of miR-138 in tissues and cell lines were determined by real-time quantitative PCR. miR-138 mimics were transfected into A549 and Calu-3 cells in vitro, and then the effects of miR-138 on lung cancer cell proliferation, cell cycle, invasion and metastasis were investigated by CCK-8 assay, Transwell and flow cytometry, respectively. The protein expression of the potential target gene Sirt1 in lung cancer cells were determined by western blot analysis. Dual-Luciferase reporter assay was performed to further confirm whether Sirt1 was the target gene of miR-138. The expression of miR-138 was significantly lower in lung cancer tissues and was negatively correlated to the differentiation degree and lymph node metastasis of lung cancer. In vitro experiment results showed that miR-138 inhibited lung cancer cell proliferation, invasion and migration. It was verified that miR-138 could downregulate Sirt1 protein expression, inhibit epithelial-mesenchymal transition (EMT), decrease the activity of AMPK signaling pathway and elevate mTOR phosphorylation level. Dual-Luciferase reporter assay demonstrated that miR-138 could directly regulate Sirt1. Downregulation of Sirt1 alone can also cause the same molecular and biological function changes. Western blot analysis and confocal microscopy results indicated that overexpression of miR-138 or interference of Sirt1 expression could inhibit lung cancer cell autophagy activity possibly through AMPK-mTOR signaling pathway. miR-138 plays a tumor suppressor function in lung cancer. It may inhibit the proliferation, invasion and migration of lung cancer through downregulation of Sirt1 expression and activation of cell autophagy. The downregulation of miR-138 is closely related to the development of lung cancer.