Chen Zehong, Wu Jialin, Huang Wensheng, Peng Jianjun, Ye Jinning, Yang Liang, Yuan YuJie, Chen Chuangqi, Zhang Changhua, Cai Shirong, He Yulong, Wu Suijing, Song Wu
Department of Gastrointestinal Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
Department of Hematology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China.
Cell Physiol Biochem. 2018;47(6):2432-2444. doi: 10.1159/000491617. Epub 2018 Jul 10.
BACKGROUND/AIMS: Gastric cancer (GC) is a common malignancy with a global incidence that ranks fourth among all tumor types. Epithelial-to-mesenchymal transition (EMT) is a tumor biological process with a role in GC cell metastasis. Long non-coding RNAs (lncRNAs) and microRNAs possess important regulatory functions at the cellular level and in diverse pathophysiological processes. This study was conducted to investigate whether lncRNA RP11-789C1.1 regulates EMT in GC by mediating the miR-5003/E-cadherin pathway.
RP11-789C1.1 and miR-5003 expression was detected in GC specimens and cell lines by quantitative real-time PCR. Western blotting and immunohistochemistry were performed to detect EMT markers in GC. Cell Counting Kit 8 assays were carried out to explore cell proliferation. Wound healing and Transwell assays were conducted to determine the migration and invasion of GC cells. To clarify the correlation between RP11-789C1.1, miR-5003, and E-cadherin, dual-luciferase reporter assays were applied.
LncRNA RP11-789C1.1 was significantly down-regulated in GC patients and cell lines, along with the concomitant up-regulation of miR-5003. Silencing RP11-789C1.1 and over-expressing miR-5003 significantly promoted the tumor behavior of GC cells. Dual-luciferase reporter assays confirmed that miR-5003 was the target of both RP11-789C1.1 and E-cadherin. Furthermore, at both the mRNA and protein level, silencing RP11-789C1.1 remarkably reduced the expression of E-cadherin and promoted EMT, which were reversed by knocking down miR-5003.
LncRNA RP11-789C1.1 inhibited EMT in GC through the RP11-789C1.1/miR-5003/E-cadherin axis, which could be a promising therapeutic target for GC.
背景/目的：胃癌（GC）是一种常见的恶性肿瘤，其全球发病率在所有肿瘤类型中排名第四。上皮-间质转化（EMT）是一种肿瘤生物学过程，在GC细胞转移中起作用。长链非编码RNA（lncRNA）和微小RNA在细胞水平和多种病理生理过程中具有重要的调节功能。本研究旨在探讨lncRNA RP11-789C1.1是否通过介导miR-5003/E-钙黏蛋白途径调节GC中的EMT。
通过定量实时PCR检测GC标本和细胞系中RP11-789C1.1和miR-5003的表达。进行蛋白质印迹和免疫组织化学检测GC中的EMT标志物。进行细胞计数试剂盒8检测以探索细胞增殖。进行伤口愈合和Transwell检测以确定GC细胞的迁移和侵袭。为了阐明RP11-789C1.1、miR-5003和E-钙黏蛋白之间的相关性，应用了双荧光素酶报告基因检测。
lncRNA RP11-789C1.1在GC患者和细胞系中显著下调，同时miR-5003上调。沉默RP11-789C1.1和过表达miR-5003显著促进了GC细胞的肿瘤行为。双荧光素酶报告基因检测证实miR-5003是RP11-789C1.1和E-钙黏蛋白的靶点。此外，在mRNA和蛋白质水平上，沉默RP11-789C1.1显著降低了E-钙黏蛋白的表达并促进了EMT，而敲低miR-5003可逆转这些变化。
lncRNA RP11-789C1.1通过RP11-789C1.1/miR-500/E-钙黏蛋白轴抑制GC中的EMT，这可能是GC的一个有前景的治疗靶点。
