miR-1297靶向HMGA1基因以调节胃癌细胞的侵裘和迁移

Effect of miR-1297 on Regulating Invasion and Migration of Gastric Cancer Cells via HMGA1 Gene

目的:探讨miR-1297是否可以通过靶向高迁移率组A1(HMGA1)调控胃癌的迁移和侵袭。方法:通过逆转录定量聚合酶链反应(RT-qPCR)测定了胃癌组织(n=30)和邻近正常组织中miR-1297和HMGA1的表达水平。体外培养胃腺癌SGC-7901细胞系和人正常胃粘膜上皮细胞GES-1。将SGC-7901细胞分为空白组、阴性对照组、miR-1297 mimics组、miR-1297 inhibitor和miR-1297 inhibitor+HMGA1 siRNA组。采用Transwell法检测SGC-7901细胞的迁移和侵袭情况。Western blot检测蛋白水平的变化。荧光素酶报告基因法检测miR-1297特异性靶基因。结果:miR-1297在胃癌组织和SGC-7901细胞中表达下调,HMGA1 mRNA水平同时升高(P<0.001),并且miR-1297与HMGA1 mRNA表达均呈负相关关系(P<0.05)。与空白组和阴性对照组相比,miR-1297 mimics组可抑制细胞在体外的迁移和侵袭能力(P<0.05)。此外,miR-1297通过靶向HMGA1的3'-UTR下调HMGA1。与空白和阴性对照相比,转染miR-1297 inhibitor下调miR-1297表达后细胞迁移和侵袭能力显著增加(P<0.05),同时转染HMGA1 siRNA(miR-1297 inhibitor+HMGA1 siRNA组)后这种促进能力被逆转(P<0.05)。结论:miR-1297通过靶向HMGA1抑制SGC-7901细胞的迁移和侵袭,提示miR-1297/HMGA1轴为胃癌提供了一种新的前瞻性治疗策略。

Objective:To investigate whether miR-1297 regulates the migration and invasion of gastric cancer by targeting high mobility group A1(HMGA1).Methods:The expression levels of miR-1297 and HMGA1 in gastric cancer(n=30)and adjacent normal tissues were determined by RT-qPCR.Gastric adenocarcinoma SGC-7901 cell line and human normal gastric mucosal epithelial cells GES-1 were cultured in vitro.SGC-7901 cells were divided into blank group,negative control group,miR-1297 mimics group,miR-1297 inhibitor and miR-1297 inhibitor+HMGA1 siRNA group.Transwell method was used to detect the migration and invasion of SGC-7901 cells.Western blot tests for changes in protein levels.The specific target genes of miR-1297 were detected by luciferase reporter gene assay.Results:The expression of miR-1297 was down-regulated in gastric cancer tissues and SGC-7901 cells,and HMGA1 mRNA level was increased at the same time(P<0.001),and miR-1297 and HMGA1 mRNA expression were negatively correlated(P<0.05).Compared with blank group and negative control group,miR-1297 mimics group could inhibit the migration and invasion ability of cells in vitro(P<0.05).In addition,miR-1297 down-regulates HMGA1 by targeting its 3'-UTR.Compared with blank and negative controls,transfection with miR-1297 inhibitor after downregulation of miR-1297 expression significantly increased cell migration and invasion ability(P<0.05).At the same time,the promoting ability was reversed after transfection of HMGA1 siRNA(miR-1297 inhibitor+HMGA1 siRNA group)(P<0.05).Conclusion:miR-1297 inhibits the migration and invasion of SGC-7901 cells by targeting HMGA1,suggesting that miR-1297/HMGA1 axis provides a new prospective therapeutic strategy for gastric cancer.