miR-9-5p靶向FGF9抑制胃癌细胞迁移侵袭的作用机制

The effect of miR-9-5p on the migration and invasion of gastric cancer cells by targeting FGF9

目的:探讨miR-9-5p在胃癌组织中的表达及对胃癌细胞株SGC7901、AGS细胞迁移和侵袭影响的作用机制。方法:real-time PCR和Western blot检测癌旁组织、胃癌组织及其细胞系中miR-9-5p和FGF9的表达;Transwell小室检测过表达miR-9-5p或敲除FGF9对SGC7901和AGS细胞迁移和侵袭能力的影响;TargetScan在线分析、双荧光素酶报告基因和Western blot实验验证miR-9-5p和FGF9的靶向关系;将转染后的SGC7901细胞分为对照(NC)组和实验(miR-9-5p)组接种于裸鼠右侧腋窝皮下,观察记录裸鼠成瘤情况并绘制生长曲线。结果:与癌旁组织相比,胃癌组织及其细胞系中miR-9-5p表达下调、FGF9表达上调(P<0.05);过表达miR-9-5p或敲除FGF9能抑制SGC7901、AGS细胞迁移和侵袭;TargetScan在线分析、双荧光素酶报告基因和Western blot实验表明miR-9-5p能调控FGF9表达;miR-9-5p组细胞成瘤速度较NC组慢(P<0.05),肿瘤体积及重量小(P<0.05)。结论:miR-9-5p在胃癌组织中表达下调,过表达miR-9-5p可抑制FGF9表达,从而抑制胃癌细胞的迁移和侵袭,减缓肿瘤生长。

Objective:To investigate the expression of miR-9-5p and FGF9 in gastric cancer and the effect of migration and invasion of gastric cancer cells SGC7901 and AGS.Methods:The expression of miR-9-5p and FGF9 in gastric cancer tissues,adjacent tissues and gastric cancer cells were detected by real-time PCR and Western blot.The migration and invasion of SGC7901 and AGS cells after overexpression of miR-9-5p or knockdown of FGF9 were detected by Transwell.The regulating relationship between miR-9-5p and FGF9 was verified by TargetScan online prediction,dual luciferase reporter system and Western blot.The transfection of SGC7901 cells,which were divided into control(NC)group and experimental(miR-9-5p)group,were inoculated into the right axilla of nude mice.The tumor formation and drawning growth curve were observed.Results:The expression of miR-9-5p in gastric cancer tissues and cell lines was significantly lower than that in gastric cancer tissues while FGF9 had the opposite trend in the tissues(P<0.05).The migration and invasion of SGC7901 and AGS cells were inhibited after overexpression of miR-9-5p or knockdown of FGF9.TargetScan online analysis,dual luciferase assay and Western blot showed that miR-9-5p negatively regulated the expression of FGF9.Compared with the NC group,the tumor formation rate of miR-9-5p group was slow(P<0.05),and the volume and weight of tumor was small(P<0.05).Conclusion:miR-9-5p is downregulated in gastric cancer tissues and overexpression of miR-9-5p can inhibit the migration and invasion of gastric cancer cells SGC7901 and AGS by directly targeting FGF9.