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LINC00665通过激活WNT-CTNNB1/β-catenin信号通路促进HeLa细胞增殖、迁移、侵袭和上皮-间质转化

夏璐^1,*, 陈毅星¹, 连加辨²

¹厦门大学附属第一医院肿瘤中心实验室，厦门 361000；²厦门大学附属第一医院检验科，厦门 361000

摘要

越来越多的证据表明，长链非编码RNA(long non-coding RNA, lncRNA)在癌症进展中起关键作用。但是，人们对lncRNA 00665 (LINC00665)在大多数癌症中的作用了解甚少。本研究旨在揭示LINC00665在宫颈癌细胞中的功能。用小发夹RNA (short hairpin RNA, shRNA)敲减HeLa细胞中LINC00665的表达以研究宫颈癌细胞在体外和体内的转移和增殖表型。将LINC00665敲减组和对照组HeLa细胞进行转录组测序，筛选差异表达基因(differentially expressed genes, DEGs)。对DEGs进行Metascape数据库功能分析和基因集富集分析(gene set enrichment analysis, GSEA)。用Western blot和免疫荧光染色法检测WNT-CTNNB1/β-catenin通路和上皮-间质转化(epithelial-mesenchymal transition, EMT)相关标记物的表达水平。结果显示，沉默LINC00665降低HeLa细胞活力，上调E-cadherin蛋白表达水平，下调N-cadherin、Vimentin和CTNNB1蛋白表达水平，抑制HeLa细胞的迁移和侵袭。生物信息学分析结果显示LINC00665可能通过激活WNT-CTNNB1/β‑catenin信号转导通路促进EMT。以上结果提示，LINC00665通过WNT-CTNNB1/β-catenin信号传导途径调控HeLa细胞增殖、迁移、侵袭和EMT。LINC00665可能作为一个潜在的宫颈癌药物治疗靶点。

关键词： 宫颈癌; 增殖; 上皮-间质转化; 长链非编码RNA; WNT信号通路

分类号：R73

LINC00665 promotes HeLa cell proliferation, migration, invasion and epithelial-mesenchymal transition by activating the WNT-CTNNB1/β‑catenin signaling pathway

XIA Lu^1,*, CHEN Yi-Xing¹, LIAN Jia-Bian²

¹Laboratory of Cancer Center, , The First Affiliated Hospital of Xiamen University, Xiamen 361000, China；²Department of Clinical Laboratory, The First Affiliated Hospital of Xiamen University, Xiamen 361000, China

Abstract

There is increasing evidence that long non-coding RNA (lncRNA) plays critical roles in cancer progression. However, the role of long non-coding RNA 00665 (LINC00665) in most cancers is poorly understood. The purpose of the present study was to reveal the functional role of LINC00665 in cervical cancer cells. HeLa cells were subjected to LINC00665 short hairpin RNA (shRNA) or control shRNA treatment to investigate the metastasis and proliferation phenotype of cervical cancer cells in vitro and in vivo. Transcriptome sequencing experiments of HeLa cells in LINC00665 silencing or control group were conducted, and the differentially 

expressed genes (DEGs) were screened. The DEGs were subjected to Metascape database functional analysis and gene set enrichment analysis. Epithelial-mesenchymal transition (EMT) related markers and a key element of WNT/β‑catenin pathway, CTNNB1 (catenin beta 1), were detected by Western blot and immunofluorescence assay. The results showed that silencing LINC00665 reduced cell viability of Hela cells, up-regulated protein expression level of E-cadherin, down-regulated protein expression levels of N-cadherin, Vimentin and CTNNB1, and inhibited cell migration and invasion of HeLa cells. Bioinformatics analysis results showed that LINC00665 might promote EMT by activating WNT-CTNNB1/β‑catenin signaling pathway. These results indicate that LINC00665 has functions in transcriptional EMT regulation via WNT-CTNNB1/β‑catenin signaling pathway and therefore can be developed as a therapeutic target for cervical cancer.

Key words: cervical cancer; proliferation; epithelial-mesenchymal transition; long noncoding RNA; WNT signaling pathway

收稿日期：2020-07-06　　录用日期：2020-09-21

通讯作者：夏璐　　E-mail: lianjiabian@163.com

DOI: 10.13294/j.aps.2021.0025

引用本文：

夏璐, 陈毅星, 连加辨. LINC00665通过激活WNT-CTNNB1/β-catenin信号通路促进HeLa细胞增殖、迁移、侵袭和上皮-间质转化[J]. 生理学报 2021; 73 (2): 233-243.

XIA Lu, CHEN Yi-Xing, LIAN Jia-Bian. LINC00665 promotes HeLa cell proliferation, migration, invasion and epithelial-mesenchymal transition by activating the WNT-CTNNB1/β‑catenin signaling pathway. Acta Physiol Sin 2021; 73 (2): 233-243