miR-183-5p通过靶向调控FAT1抑制胃癌进展

miR-183-5p Inhibits Progression of Gastric Cancer by Targeting FAT1

目的探讨microRNA-183-5p(miR-183-5p)在胃癌进展中的作用及其机制。方法收集2018年5月至2020年5月河北大学附属医院66例胃癌(GC)患者临床标本资料,采用生物信息学工具TargetScan和miRanda预测miR-183-5p可能的靶标,采用Western blot法和定量聚合酶链反应检测GC肿瘤组织和细胞系中miR-183-5p、非典型钙粘蛋白1(FAT1)水平。采用荧光素酶法验证miR-183-5p和FAT1之间的相互作用。结果GC患者肿瘤组织和GC细胞株中miR-183-5p水平显著降低。GC肿瘤组织miR-183-5p与肿瘤大小、肿瘤分型、TNM分期及患者预后(生存时间)呈负相关。体外实验发现,GC细胞株BGC-823过表达miR-183-5p后降低了其FAT1的表达,抑制细胞增殖并促进细胞凋亡。荧光素酶报告基因实验确认FAT1是miR-183-5p靶向基因。GC肿瘤组织miR-183-5p和FAT1水平呈现出明显负相关。将miR-183-5p转染入GC细胞BGC-823内能够抑制其在移植瘤模型中的生长。结论miR-183-5p潜在靶标是FAT1,miR-183-5p通过抑制FAT1负性调控GC的发展,miR-183-5p有望成为GC潜在的诊断和治疗靶点。

Objective To investigate potential role of microRNA-183-5p(miR-183-5p)in the progress of gastric cancer(GC),and to identify its potential mechanism.Methods A total of 66 clinical specimens from GC patients admitted in Affiliated Hospital of Hebei University from May 2018 to May 2020 were collected.The possible targets of miR-183-5p were predicted by using bioinformatics prediction tools TargetScan and miRanda.Western blotting and quantitative polymerase chain reaction were used to investigate miR-183-5p and FAT atypical cadherin 1(FAT1)levels in tissues and cell lines of GC.Luciferase assay was employed to verify predicted interaction between miR-183-5p and FAT1.Results miR-183-5p was decreased in GC tissues and cell lines compared with control subjects,and was negatively correlated with tumor size,tumor type,TNM staging and overall survival.Transfection of miR-183-5p downregulated FAT1 expression,reduced cell proliferation and promoted apoptosis of BGC-823 in vitro.Luciferase assay confirmed that FAT1 was targeted by miR-183-5p.A significant negative correlation was observed between miR-183-5p and FAT1 expression in GC tissues.Furthermore,upregulation of miR-183-5p inhibited the tumor growth by decreasing FAT1 in vivo.Conclusion TargetScan and miRanda predict that FAT1 is a potential target of miR-183-5p.miR-183-5p negatively mediates GC development through suppressing FAT1,and might be a promising target for diagnosis and treatment of GC in the future.