微小RNA-144对人结肠癌细胞株增殖、侵袭迁移影响及其与同源盒基因A10结合力  

Effects of microRNA-144 on proliferation,invasion,and migration of human colon cancer cell lines and its binding ability to homeobox gene A10

作　　者：李萍[1] 张月晓[1] 符皓 时军利[1] 李炳庆[1] LI Ping;ZHANG Yuexiao;FU Hao;SHI Juni;LI Bingqing(Department of Gastroenterology,Affiliated Hospital of Chengde Medical College,Chengde 067000,China;不详)

机构地区：[1]承德医学院附属医院消化内科,河北承德067000 [2]承德医学院附属医院营养科

出　　处：《山东医药》2023年第30期16-20,共5页Shandong Medical Journal

基　　金：承德市科技支撑计划项目(202006A077)。

摘　　要：目的 观察微小RNA-144(miR-144)对人结肠癌细胞株增殖、迁移、侵袭的影响及其与同源盒基因A10(homeobox gene A10,HOXA10)的结合力。方法 (1)人结肠癌细胞株HCT116、SW480、CL-11和永生化正常人结肠上皮细胞株FHC中miR-144及HOXA10 mRNA检测:采用实时荧光定量PCR法检测HCT116、SW480、CL-11及FHC中miR-144与HOXA10 mRNA,观察人结肠癌细胞和人结肠上皮细胞miR-144、HOXA10 mRNA表达变化,选择miR-144相对表达量最低HCT116细胞为后续研究对象。(2)miR-144对人结肠癌细胞株增殖、侵袭及迁移的影响观察:取对数生长期HCT116细胞分为3组,1组细胞顺序转染miR-144模拟物(miR-144 mimics)和HOXA10过表达质粒pcDNA3.1-HOXA10,2组细胞转染miR-144 mimics,3组细胞转染空白质粒。转染24 h时测算各组细胞增殖活性、观察各组细胞的侵袭及迁移情况、检测各组细胞miR-144及HOXA10蛋白。(3)HCT116细胞中miR-144与HOXA10的结合力观察:取对数生长期HCT116细胞分为4组:A组先后转染野生型HOXA10荧光素酶报告基因质粒(HOXA10-WT)、miR-144mimics,B组先后转染HOXA10-WT、空白对照NC-mimics,C组先后转染突变型HOXA10-MUT、miR-144 mimics,D组先后转染HOXA10-MUT、NC-mimics,转染24 h时采用双荧光素酶检测试剂测算各组细胞的相对荧光素酶活性(代表HOXA10、miR-144的靶向结合力)。结果 与FHC细胞比较,HCT116、SW480、CL-11细胞miR-144相对表达量低,HOXA10 mRNA相对表达量高(P均<0.05)。与3组比较,2组细胞miR-144相对表达量高(P<0.05),培养24、48、72 h时细胞OD值均降低,迁移及侵袭穿膜细胞数少,HOXA10蛋白相对表达量低(P均<0.05);与2组比较,1组细胞HOXA10蛋白相对表达量高,培养24、48、72 h时细胞的OD值均高,迁移及侵袭穿膜细胞数多。与B组比较,A组HCT116细胞的相对荧光素酶活性低(P<0.05)。结论 miR-144可抑制结肠癌细胞的增殖、侵袭及迁移,同时抑制细胞HOXA10蛋白表达。HCT116细胞中miR-144与HOXA10靶向结Objective To observe the effects of microRNA-144(miR-144) on the proliferation,migration,and invasion of human colon cancer cell lines and its binding ability to homeobox gene A10(HOXA10).Methods(1) Detection of miR-144 and HOXA10 mRNA in human colon cancer cell lines HCT116,SW480,CL-11,and immortalized normal human colon epithelial cell line FHC:Real-time fluorescence quantitative PCR was used to detect miR-144 and HOXA10 mRNA in HCT116,SW480,CL-11,and FHC,and we observed the changes in miR-144 and HOXA10 mRNA expression in human colon cancer cells and human colon epithelial cells.HCT116 cells with the lowest relative expression of miR-144 were selected as the follow-up study subjects.(2) Observation of the effects of miR-144 on the proliferation,invasion,and migration of human colon cancer cell lines:HCT116 cells in the logarithmic growth phase were divided into three groups.Cells in the Group 1 were sequentially transfected with miR-144 mimics and HOXA10 overexpression plasmid pcDNA3.1-HOXA10.Cells in the Group 2 were transfected with miR-144 mimics,and cells in the Group 3 were transfected with blank plasmids.At 24 h of transfection,the proliferation activity of cells,and the cell invasion and migration in each group were measured,and the miR-144 and HOXA10 proteins of each group were detected.(3) Observation on the binding ability of miR-144 and HOXA10 in HCT116 cells:HCT116 cells in the logarithmic growth phase were divided into four groups:Cells in the Group A were transfected with wild-type HOXA10 luciferase reporter gene plasmid(HOXA10-WT) and miR-144 mimics,cells in the Group B were transfected with HOXA10-WT and blank control NC-mimics,cells in the Group C were transfected with mutant HOXA10-MUT and miR-144 mimics,and cells in the Group D were transfected with HOXA10-MUT and NC-mimics.At 24 h of transfection,dual luciferase detection reagents were used to measure the relative luciferase activity of cells in each group(representing the targeted binding force of HOXA10 and miR-144).Results Compared with

关 键 词：微小RNA 微小RNA-144 同源盒基因A10 结肠癌 细胞增殖 细胞迁移 细胞侵袭 

分 类 号：R656.9[医药卫生—外科学]