环状RNA ATP2B1靶向miR-452-5p降低胃癌细胞系的增殖和侵袭  

Circular RNA ATP2B1 reduces proliferation and invasion of gastric cancer cell lines by targeting miR-452-5p

作　　者：郑彬彬 林淑莹[1] 李德隆 陈杰 黄子成[1] ZHENG Binbin;LIN Shuying;LI Delong;CHEN Jie;HUANG Zicheng(Department of Gastroenterology,Quanzhou First Hospital Affiliated to Fujian Medical University,Quanzhou 476100;Department of Gastroenterology,Jinshan Hospital,Fudan University,Shanghai 200540,China)

机构地区：[1]福建医科大学附属泉州第一医院胃肠消化内科,福建泉州476100 [2]复旦大学附属金山医院消化内科,上海200540

出　　处：《基础医学与临床》2023年第6期882-888,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金(81702356)。

摘　　要：目的探讨环状RNA circATP2B1对胃癌细胞增殖和侵袭的影响及其可能的分子机制。方法收集2018年7月至2021年2月福建医科大学附属泉州第一医院胃肠肝胆外科行手术切除并病理学诊断的44例胃癌组织标本及癌旁组织标本。选取4株胃癌细胞系(SGC7901、HS-746T、MGC803、BGC823)和正常胃黏膜上皮细胞系(GES-1)。RT-qPCR检测胃癌组织和细胞系中circATP2B1表达。将circATP2B1表达最低的胃癌细胞分为对照组(转染阴性对照质粒)和实验组(转染circATP2B1过表达质粒)。分别采用MTT法和Transwell小室法检测各组胃癌细胞的增殖活性和侵袭能力。生物信息学和双荧光素酶报告基因实验分析circATP2B1可能的作用机制,RT-qPCR和Western blot检测circATP2B1下游基因的表达。结果胃癌组织circATP2B1表达量显著低于癌旁组织(0.92±0.08 vs 3.62±0.23,P<0.01)。胃癌细胞系circATP2B1表达量均显著低于GES-1细胞(P<0.01),其中以HS-746T细胞的表达量最低(P<0.01)。与对照组相比,实验组HS-746T细胞的增殖活性显著降低(P<0.05),侵袭能力显著下降(P<0.01)。生物信息学和双荧光素酶报告基因实验显示circATP2B1可靶向结合miR-452-5p(P<0.01),miR-452-5p可靶向结合原钙黏附蛋白9(PCDH9)(P<0.01)。与对照组比较,实验组HS-746T细胞miR-452-5p表达显著下降(1.00±0.04 vs 0.24±0.05,P<0.01),PCDH9基因表达显著上升(P<0.01)。结论circATP2B1在胃癌组织和细胞系中低表达,circATP2B1通过靶向结合miR-452-5p正调控PCDH9基因表达,进而降低胃癌HS-746T细胞增殖和侵袭能力。Objective To explore the effects of circular RNA circATP2B1 on proliferation and invasion of gastric cancer cells and its potential molecular mechanism.Methods From July 2018 to February 2021,44 cases of gastric cancer tissue specimens and paracancerous tissue specimens were collected from the Department of Gastrointestinal Hepatobiliary Surgery of Quanzhou First Hospital Affiliated to Fujian Medical University.Four gastric cancer cell lines(SGC7901,HS-746T,MGC803,BGC823)and normal gastric mucosal epithelial cells(GES-1)were selected.The expression of circATP2B1 in gastric cancer tissues and cell lines was examined by RT-PCR.The gastric cancer cells with the lowest expression of circATP2B1 were divided into control group(transfected with negative control plasmid)and experimental group(transfected with circATP2B1 over expression plasmid).MTT and Transwell experiment were used to detect the proliferation activity and invasion ability of gastric cancer cells in each group.Bioinformatics and dual luciferase reporter gene experiments were used to analyze the possible molecular mechanism of circATP2B1,and RT-qPCR and Western blot were used to detect the expression of circATP2B1 downstream genes.Results The expression of circATP2B1 in gastric cancer tissue was significantly lower than that in adjacent tissues(0.92±0.08 vs.3.62±0.23,P<0.01).The expression of circATP2B1 in gastric cancer cell lines was significantly lower than that in GES-1 cells(P<0.01)and HS-746T cells had the lowest expression level(P<0.01).Compared with the control group,the proliferation activity of HS-746T cells in the experimental group was significantly reduced(P<0.05),and the invasion ability was significantly reduced(P<0.01).Bioinformatics and dual luciferase reporter gene experiments showed that circATP2B1 can target miR-452-5p(P<0.01),and miR-452-5p can target protocadherin 9(PCDH9)(P<0.01).Compared with the control group,the expression of miR-452-5p in HS-746T cells in the experimental group decreased significantly(1.00±0.04 vs.0.24±0.05,

关 键 词：胃癌 环状RNA circATP2B1 miR-452-5p 原钙黏附蛋白9 

分 类 号：R735.1[医药卫生—肿瘤]