lncRNA XIST通过靶向miR-3666调控胃癌细胞增殖、侵袭转移机制

lncRNA XIST regulates proliferation,invasion and metastasis of gastric cancer cells by targeting miR-3666

目的:探讨lncRNA XIST对胃癌细胞增殖、侵袭转移的影响及分子机制。方法:收集武汉市第四医院30例胃癌患者癌组织及癌旁组织,培养正常胃黏膜上皮细胞MRG-1、胃癌细胞SGC-7901和AGS,采用实时荧光定量PCR(RT-qPCR)检测lncRNA XIST和miR-3666表达水平;通过敲减胃腺癌细胞AGS中lncRNA XIST表达以及下调miR-3666表达水平,分析lncRNA XIST及miR-3666对胃癌细胞的调控作用,将细胞AGS分为si-NC组、si-XIST组、miR-NC组、miR-3666组、si-XIST+antimiR-NC组、si-XIST+anti-miR-3666组;MTT检测细胞增殖情况;Transwell检测细胞迁移和侵袭;流式细胞术检测细胞凋亡;双荧光素酶报告实验检测lncRNA XIST和miR-3666的靶向关系。结果:与癌旁组织和正常胃黏膜上皮细胞MRG-1比较,胃癌组织和AGS、SGC-7901细胞中XIST表达水平升高,miR-3666表达水平降低(P<0.05)。敲除lncRNA XIST或过表达miR-3666,AGS细胞增殖能力减弱,迁移和侵袭能力降低,细胞凋亡率升高(P<0.05)。lncRNA XIST靶向调控miR-3666,抑制miR-3666逆转了XIST基因敲除对胃癌AGS细胞增殖、运动性及凋亡的影响。结论:敲除lncRNA XIST可通过上调miR-3666抑制胃癌细胞增殖、迁移和侵袭,并促进细胞凋亡。

Objective:To investigate the effect of lncRNA XIST on the proliferation,invasion and metastasis of gastric cancer cells and its molecular mechanism.Methods:The cancer tissues and adjacent tissues of 30 gastric cancer patients in Wuhan No.4Hospital were collected,cultured normal gastric mucosal epithelial cells MRG-1,gastric cancer cells SGC-7901 and AGS,and used real-time fluorescent quantitative PCR(RT-qPCR)to detect the expression of lncRNA XIST and miR-3666;by knocking down the expression of lncRNA XIST in gastric adenocarcinoma cells AGS and down-regulating the expression level of miR-3666,the regulation of lncRNA XIST and miR-3666 on gastric cancer cells was analyzed,and the cells AGS were divided into si-NC group,si-XIST group,miR-NC group,miR-3666 group,si-XIST+anti-miR-NC group,si-XIST+anti-miR-3666 group;MTT was used to detect cell proliferation;Transwell was used to detect cell migration and invasion;flow cytometry was used to detect the cell apoptosis;dual luciferase reporter experiment was used to detect the targeting relationship between lncRNA XIST and miR-3666.Results:Compared with adjacent tissues and MRG-1 cells,expression of XIST in gastric cancer tissue,AGS and SGC-7901 cells were increased,and the expression of miR-3666 was decreased(P<0.05).Knockout lncRNA XIST or overexpress miR-3666,AGS cells proliferation ability was weakened,the migration and invasion ability were reduced,and the cell apoptosis rate was increased(P<0.05).lncRNA XIST targeted and regulated miR-3666;inhibited miR-3666 reversed the effect of XIST gene knockout on the proliferation,motility and apoptosis of gastric cancer AGS cells.Conclusion:Knockout of lncRNA XIST can inhibit the proliferation,migration and invasion of gastric cancer cells by up-regulating miR-3666,and promote cell apoptosis.