circFOXM1作为新型胃癌标志物调控胃癌发生、发展的机制研究  

Study on the mechanism of circFOXM1 as a new gastric cancer marker regulating the occurrence and development of gastric cancer

作　　者：朱沙沙[1] 马小宏 程莉莉 谢逸尘 ZHU Shasha;MA Xiaohong;CHENG Lili;XIE Yichen(Department of Clinical Laboratory,Rugao People′s Hospital,Rugao,Jiangsu 226500,China)

机构地区：[1]江苏省如皋市人民医院检验科,江苏如皋226500

出　　处：《检验医学与临床》2024年第11期1586-1594,共9页Laboratory Medicine and Clinic

基　　金：2021年江苏省如皋市指导性科技攻关计划项目(RG2021SZD005)。

摘　　要：目的分析新型胃癌标志物环状叉头框蛋白M1(circFOXM1)在胃癌发生、发展机制中的调控作用。方法选取2022年1月至2023年4月该院肿瘤科收治的60例胃癌患者作为研究对象,检测其胃癌组织及癌旁组织中circFOXM1的表达水平。根据circFOXM1的表达水平,将60例患者分为circFOXM1高表达组和circFOXM1低表达组。比较circFOXM1高表达组和circFOXM1低表达组的临床病理资料(淋巴结转移、肿瘤侵袭深度和临床分期)。采用实时荧光定量聚合酶链反应(PCR)和免疫印迹法检测GES-1、HGC-27、BGC-823、MKN-45、MKN-1细胞中circFOXM1信使RNA(mRNA)、微小RNA(miR)-182-5p mRNA与母亲信号蛋白同源物7(SMAD7)mRNA及SMAD7蛋白的表达水平。体外培养MKN-45细胞,将其随机分为对照组、circFOXM1敲低组[转染circFOXM1小干扰RNA(siRNA)质粒]、circFOXM1过表达组(转染circFOXM1过表达质粒)、共转染阴性对照组(转染空载质粒和miR-182-5p抑制剂阴性对照)、circFOXM1敲低+miR-182-5p抑制剂组(转染circFOXM1 siRNA质粒和miR-182-5p抑制剂),分组转染后,采用实时荧光定量PCR和免疫印迹法检测各组MKN-45细胞circFOXM1 mRNA、miR-182-5p mRNA与SMAD7 mRNA及SMAD7蛋白的表达水平;采用Transwell侵袭及细胞划痕试验检测各组MKN-45细胞侵袭迁移情况;采用四甲基偶氮唑蓝(MTT)法检测各组MKN-45细胞增殖情况。取50只裸鼠随机分为对照小鼠组、circFOXM1敲低小鼠组、circFOXM1过表达小鼠组、共转染阴性对照小鼠组、circFOXM1敲低+miR-182-5p抑制剂小鼠组,每组10只。将各组MKN-45细胞接种在对应裸鼠右腋附近背部皮下构建胃癌移植瘤模型,饲养4周后检测各组移植瘤裸鼠MKN-45细胞增殖率、肿瘤体积及肿瘤质量。将MKN-45细胞随机分为野生miR-182-5p+空载组、野生miR-182-5p+circFOXM1过表达组、突变miR-182-5p+空载组、突变miR-182-5p+circFOXM1过表达组、野生SMAD7+miR-182-5p mimic阴性对照组、野生SMAD7+miR-182-5p mimic组、Objective To analyze the regulatory role of a new gastric cancer marker circular forkhead box M1(circFOXM1)in the occurrence and development of gastric cancer.Methods Sixty patients with gastric cancer admitted to the Oncology Department of the hospital from January 2022 to April 2023 were selected as the research objects.The expression level of circFOXM1 in gastric cancer tissues and adjacent tissues was detected.Sixty patients were divided into circFOXM1 high expression group and circFOXM1 low expression group.The clinicopathological data(lymph node metastasis,depth of tumor invasion and clinical stage)of circFOXM1 high expression group and circFOXM1 low expression group were compared.Real-time fluorescent quantitative polymerase chain reaction(PCR)and Western blot were used to detect circFOXM1 massage RNA(mRNA)and microRNA(miR)-182-5p and maternal signaling protein homolog 7(SMAD7)mRNA and SMAD7 protein expression levels in GES-1,HGC-27,BGC-823,MKN-45 and MKN-1 cells were cultured in vitro.They were randomly divided into control group,circFOXM1 knockdown group[transfected with circFOXM1 small interfering RNA(siRNA)plasmid],circFOXM1 overexpression group(transfected with circFOXM1 overexpression plasmid)and co-transfected negative control group(transfected with empty plasmid and miR-182-5p inhibitor negative control),circFOXM1 knockdown+miR-182-5p inhibitor group(transfected with circFOXM1 siRNA plasmid and miR-182-5p inhibitor).After group transfection,real-time fluorescent quantitative PCR and Western blot were used to detect the expression levels of circFOXM1 mRNA,miR-182-5p mRNA,SMAD7 mRNA and SMAD7 protein in MKN-45 cells of each group.Transwell invasion and cell scratch test were used to detect the invasion and migration of MKN-45 cells in each group.The proliferation of MKN-45 cells was detected by methyl thiazolyl tetrazolium(MTT)assay.Fifty nude mice were randomly divided into control mice group,circFOXM1 knockdown mice group,circFOXM1 overexpression mice group,co-transfection negative control mice gro

关 键 词：环状叉头框蛋白M1 微小RNA-182-5p 母亲信号蛋白同源物7 胃癌 机制 

分 类 号：R735.2[医药卫生—肿瘤] R446.8[医药卫生—临床医学]