miR⁃30a⁃5p靶向与调控SNAIL1抑制人胃癌细胞BGC⁃823的迁移  

miR-30a-5p Targets and Regulates SNAIL1 to Inhibit the Migration of Human Gastric Cancer BGC-823 cells

作　　者：李欣 张富蕊 周文淼 张乐乐 施天怡 张震[2] 刘昆梅 韩学波[1] 郭乐[1] LI Xin;ZHANG Fu-Rui;ZHOU Wen-Miao;ZHANG Le-Le;SHI Tian-Yi;ZHANG Zhen;LIU Kun-Mei;HAN Xue-Bo;GUO Le(Key Laboratory of Clinical Pathogenic Microbiology,Ningxia Medical University,Yinchuan 750004,China;Department of Gerontology and Special Needs,General Hospital of Ningxia Medical University,Yinchuan 750004,China;Key Laboratory of Craniocerebral Diseases,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学临床病原微生物重点实验室,银川750004 [2]宁夏医科大学总医院老年与特需科,银川750004 [3]宁夏医科大学颅脑疾病重点实验室,银川750004

出　　处：《中国生物化学与分子生物学报》2023年第3期438-447,共10页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金(No.82160497、No.32070930);宁夏自然科学基金(No.2022AAC02034、No.2020AAC03154);宁夏重点研发计划项目(No.2020BFG02012);宁夏高等学校科学研究项目(No.NGY2020043)资助。

摘　　要：已有研究表明,miR-30a-5p在胃癌中发挥抑癌基因的作用,抑制胃癌细胞的增殖与侵袭,但其抑制胃癌发生发展的机制尚不清楚。本文研究miR-30a-5p如何靶向调控SNAIL 1,进而影响人胃癌细胞BGC-823的迁移。通过qRT-PCR检测和TCGA数据库分析发现,miR-30a-5p的表达水平在胃癌组织与胃癌细胞BGC-823中均降低。利用划痕实验和Transwell实验检测发现,在BGC-823细胞中转染miR-30a-5p mimics过表达miR-30a-5p后细胞迁移和侵袭明显被抑制,而转染miR-30a-5p inhibitor降低miR-30a-5p表达水平时细胞迁移和侵袭能力提高。qRT-PCR和Western印迹及免疫荧光检测发现,过表达miR-30a-5p细胞中波形蛋白(vimentin)和N-钙黏着蛋白(N-cadherin)的mRNA及蛋白质表达水平降低,而E-钙黏着蛋白(E-cadherin)表达水平升高。在miR-30a-5p inhibitor组中结果恰恰相反。通过生物信息学预测、双荧光素酶基因报告、Western印迹及免疫荧光验证SNAIL 1是miR-30a-5p作用的靶基因。在过表达miR-30a-5p的BGC-823细胞中进一步过表达SNAIL 1,可以逆转miR-30a-5p的作用。该研究表明,miR-30a-5p可通过负调控靶基因SNAIL 1的表达,进而抑制BGC-823胃癌细胞的迁移。本研究将为miR-30a-5p在胃癌发生发展中的作用机制提供理论依据,并有望为胃癌治疗提供新靶点和生物标志物。Previous studies have shown that miR-30a-5p plays a role as a tumor suppressor gene in gastric cancer,inhibiting the proliferation and invasion of gastric cancer cells,but its specific mechanisms remains unclear.In this study,we study the mechanism that miR-30a-5p targets SNAIL 1 and affects the migration of human gastric cancer BGC-823 cells.Analysis results from qRT-PCR and TCGA database discovered that the expression level of miR-30a-5p was decreased in both gastric cancer tissues and BGC-823 cells.Using scratch assays and Transwell experiments,it was found that cell migration was significantly inhibited after transfection of miR-30a-5p mimics in BGC-823 cells to overexpress miR-30a-5p,while transfection of miR-30a-5p inhibitor decreased miR-30a-5p expression levels and improved cell migration ability.qRT-PCR,Western blot and immunofluorescence detection showed that the mRNA and protein expression levels of Vimentin and N-cadherin in miR-30a-5p overexpressed cells decreased,while the expression level of E-cadherin increased.The results were reversed in miR-30a-5p inhibitor group.SNAIL 1 was verified as the target gene of miR-30a-5p by bioinformatics prediction,dual luciferase reporter assay,Western blot and immunofluorescence analysis.Further overexpression of SNAIL 1 in BGC-823 cells overexpressing miR-30a-5p reversed the effect of miR-30a-5p.In this study,miR-30a-5p inhibits the migration of gastric cancer BGC-823 cells by negatively regulating the expression of target gene SNAIL 1.This study will provide theoretical basis for the mechanism of miR-30a-5p in the occurrence and development of gastric cancer,and is expected to provide new biomarkers and targets for gastric cancer.

关 键 词：胃癌 miR-30a-5p SNAIL 1 迁移 

分 类 号：R735.2[医药卫生—肿瘤]