微小RNA-183通过下调MTA1在人类非小细胞肺癌中发挥肿瘤抑制作用。
MicroRNA-183 Acts as a Tumor Suppressor in Human Non-Small Cell Lung Cancer by Down-Regulating MTA1.
作者信息
Yang Cheng-Liang, Zheng Xiao-Li, Ye Ke, Ge Hong, Sun Ya-Nan, Lu Yu-Fei, Fan Qing-Xia
机构信息
Department of Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Department of Radiation Oncology, Affiliated Cancer Hospital of Zhengzhou University, Henan Cancer Hospital, Zhengzhou, China.
出版信息
Cell Physiol Biochem. 2018;46(1):93-106. doi: 10.1159/000488412. Epub 2018 Mar 20.
BACKGROUNDS/AIMS: MicroRNAs (miRs) often contribute to the progression of non-small cell lung cancer (NSCLC) via regulation of mRNAs that are involved in lung homeostasis. We conducted a study aimed at exploring the roles of miR-183 in the proliferation, epithelial-mesenchymal transition (EMT), invasion and migration of human NSCLC cells via targeting MTA1.
METHODS
NSCLC and adjacent normal tissues were collected from 194 patients with NSCLC. Positive expression of MTA1 protein was detected by immunohistochemistry. The highest levels of expression of miR-183 were detected using RT-qPCR in SPC-A-1 cells, which were selected and assigned to the following groups: blank, negative control (NC), miR-183 mimic, miR-183 inhibitor, siRNA-MTA1, and miR-183 inhibitor + siRNA-MTA1. The expression of miR-183 and the mRNA and protein expression of MTA1, E-cadherin, Vimentin, Snail, PCNA, Bax and Bcl-2 in tissues and transfected cells were measured using RT-qPCR and western blot analysis. Cell proliferation, apoptosis, migration and invasion were evaluated by CCK-8, flow cytometry, scratch tests and Transwell assays. Tumor xenografts were conducted in nude mice to determine tumor growth.
RESULTS
SPC-A-1 cells with the highest levels of miR-183 expression were selected. Compared with adjacent normal tissues, the expression of miR-183 and the mRNA and protein expression of E-cadherin and Bax were decreased in NSCLC tissues, while mRNA and protein expression of MTA1, Vimentin, snail, PCNA and Bcl-2 were increased. MiR-183 was over-expressed in the miR-183 mimic group and under-expressed in the miR-183 inhibitor and miR-183 inhibitor + siRNA-MTA1 groups. In the miR-183 mimic and siRNA-MTA1 groups, the mRNA and protein expression of E-cadherin and Bax, as well as cell apoptosis, were enhanced, while the expression levels of MTA1, Vimentin, snail, PCNA and Bcl-2 mRNA and protein, cell proliferation, migration, invasion and tumor growth were reduced relative to the blank and NC groups. The miR-183 inhibitor group exhibited an opposite trend.
CONCLUSION
Our study indicates that miR-183 down-regulates MTA1 to inhibit the proliferation, EMT, migration and invasion of human NSCLC cells.
背景/目的：微小RNA（miR）常通过调控参与肺稳态的mRNA来促进非小细胞肺癌（NSCLC）的进展。我们开展了一项研究，旨在探讨miR-183通过靶向MTA1在人NSCLC细胞增殖、上皮-间质转化（EMT）、侵袭和迁移中的作用。
方法
收集194例NSCLC患者的NSCLC组织及癌旁正常组织。采用免疫组织化学法检测MTA1蛋白的阳性表达。在SPC-A-1细胞中，通过RT-qPCR检测到miR-183的最高表达水平，将其选出并分为以下几组：空白组、阴性对照组（NC）、miR-183模拟物组、miR-183抑制剂组、siRNA-MTA1组以及miR-183抑制剂+siRNA-MTA1组。采用RT-qPCR和蛋白质印迹分析检测组织及转染细胞中miR-183的表达以及MTA1、E-钙黏蛋白、波形蛋白、Snail、增殖细胞核抗原（PCNA）、Bax和Bcl-2的mRNA和蛋白表达。通过CCK-8法、流式细胞术、划痕试验和Transwell试验评估细胞增殖、凋亡、迁移和侵袭。在裸鼠中进行肿瘤异种移植以确定肿瘤生长情况。
结果
选择了miR-183表达水平最高的SPC-A-1细胞。与癌旁正常组织相比，NSCLC组织中miR-183的表达以及E-钙黏蛋白和Bax的mRNA和蛋白表达降低，而MTA1、波形蛋白、Snail、PCNA和Bcl-2的mRNA和蛋白表达增加。miR-183在miR-183模拟物组中过表达，在miR-183抑制剂组和miR-183抑制剂+siRNA-MTA1组中低表达。在miR-183模拟物组和siRNA-MTA1组中，E-钙黏蛋白和Bax的mRNA和蛋白表达以及细胞凋亡增强，而相对于空白组和NC组，MTA1、波形蛋白、Snail、PCNA和Bcl-2的mRNA和蛋白表达水平、细胞增殖、迁移、侵袭及肿瘤生长均降低。miR-183抑制剂组表现出相反的趋势。
结论
我们的研究表明，miR-183通过下调MTA1来抑制人NSCLC细胞的增殖、EMT、迁移和侵袭。
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