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长链非编码RNA MEG3通过调控WNT/β-连环蛋白信号通路抑制口腔鳞状细胞癌的增殖和转移。

Long non-coding RNA MEG3 inhibits the proliferation and metastasis of oral squamous cell carcinoma by regulating the WNT/β-catenin signaling pathway.

作者信息

Liu Zongxiang, Wu Cui, Xie Nina, Wang Penglai

机构信息

Department of Oral and Maxillofacial Surgery, Xuzhou Stomatological Hospital, Xuzhou, Jiangsu 221006, P.R. China.

School of Stomatology, Xuzhou Medical University, Xuzhou, Jiangsu 221004, P.R. China.

出版信息

Oncol Lett. 2017 Oct;14(4):4053-4058. doi: 10.3892/ol.2017.6682. Epub 2017 Jul 27.

Abstract

This study aimed to investigate how long non-coding RNA (lncRNA) maternally expressed gene 3 (MEG3) inhibits the growth and metastasis of oral squamous cell carcinoma (OSCC) by regulating WNT/β-catenin signaling pathway in order to explore the antitumor effect of MEG3 and to provide a potential molecular target for the treatment of OSCC. The RT-qPCR technique was used to quantitatively analyze the expression of MEG3 in cancer and adjacent tissues collected from the patients after surgery. Using the Lipofectamine method, the MEG3 overexpression vector and the siRNA interference vector were constructed and transfected into SCC15 and Cal27 cells, respectively, followed by cell proliferation, apoptosis and metastasis analyses. The semi-quantitative analysis of the expression of the β-catenin protein in transfected cells was performed by the western blot analysis, and the activity of the WNT/β-catenin signaling pathway was analyzed using the TOP/FOP flash reporters. In addition, the cells were treated with decitabine to investigate the correlation between the MEG3 expression and the DNA methylation. Results showed that the expression level of MEG3 was significantly decreased in OSCC (p<0.05) and overexpression of MEG3 inhibited the proliferation and metastasis of cancer cells and promoted apoptosis. Importantly, MEG3 played a role as a tumor suppressor by inhibiting the WNT/β-catenin signaling pathway. In addition, the expression of the MEG3 was significantly affected by the degree of DNA methylation. It was concluded that the lncRNA MEG3 can inhibit the growth and metastasis of OSCC by negatively regulating the WNT/β-catenin signaling pathway.

摘要

本研究旨在探讨长链非编码RNA(lncRNA)母系表达基因3(MEG3)如何通过调节WNT/β-连环蛋白信号通路来抑制口腔鳞状细胞癌(OSCC)的生长和转移,以探索MEG3的抗肿瘤作用,并为OSCC的治疗提供潜在的分子靶点。采用RT-qPCR技术对手术患者癌组织及癌旁组织中MEG3的表达进行定量分析。利用Lipofectamine法分别构建MEG3过表达载体和siRNA干扰载体,并转染至SCC15和Cal27细胞,随后进行细胞增殖、凋亡及转移分析。通过蛋白质免疫印迹分析对转染细胞中β-连环蛋白的表达进行半定量分析,利用TOP/FOP荧光素酶报告基因分析WNT/β-连环蛋白信号通路的活性。此外,用阿扎胞苷处理细胞,以研究MEG3表达与DNA甲基化之间的关系。结果显示,OSCC中MEG3的表达水平显著降低(p<0.05),MEG3过表达抑制癌细胞的增殖和转移并促进凋亡。重要的是,MEG3通过抑制WNT/β-连环蛋白信号通路发挥肿瘤抑制作用。此外,MEG3的表达受DNA甲基化程度的显著影响。结论是lncRNA MEG3可通过负调控WNT/β-连环蛋白信号通路抑制OSCC的生长和转移。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e539/5607653/68479713de7e/ol-14-04-4053-g00.jpg

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