【摘要】 目的:探讨miR-26a靶向高迁移率族蛋白1 (HMGA1)基因对结肠癌细胞生长、侵袭和迁移的影响,阐明HMGA1基因是否为miR-26a的靶基因。方法:将miR-NC (对照)、miR-26amimics (模拟物)和miR-26ainhibitor (抑制物)转染人结肠癌SW480细胞作为miR-NC组、miR-26amimics组和miR-26ainhibitor组。RT-PCR和Western blotting法分别检测各组SW480细胞中HMGA1mRNA和蛋白表达水平。采用荧光素酶报告基因检测试剂盒检测各组SW480细胞中双荧光素酶活性,以此确定HMGA1是否为miR-26a的靶基因。采用CCK8实验检测各组细胞增殖活力,Transwell小室法检测各组细胞侵袭数和迁移数。结果:HMGA1基因是miR-26a的靶基因。与miR-NC组比较,转染24、48和72h时miR-26amimics组SW480细胞增殖活力明显降低(P<0.01),而miR-26ainhibitor组细胞增殖活力明显升高(P<0.01)。与miR-NC组比较,各时间点miR-26amimics组和miR-26amimics+pcDNA3.1-HMGA1组细胞增殖活力、细胞侵袭数和细胞迁移数均降低(P<0.01),而miR-NC+pcDNA3.1-HMGA1组细胞增殖活力、细胞侵袭数和细胞迁移数均升高(P<0.01);与miR-26amimics组比较,各时间点miR-26amimics+pcDNA3.1-HMGA1组细胞增殖活力、细胞侵袭数和细胞迁移数均升高(P<0.01);与miR-NC+pcDNA3.1-HMGA1组比较,各时间点miR-26a+pcDNA3.1-HMGA1组细胞增殖活力、细胞侵袭数和细胞迁移数均降低(P<0.01)。结论:HMGA1是miR-26a的靶基因。上调miR-26a表达可抑制人结肠癌SW480细胞增殖,下调miR-26a表达可促进人结肠癌SW480细胞增殖。更多还原

【Abstract】 Objective:To investigate the effects of miR-26 atargeting high mobility group protein 1(HMGA1)gene on the growth,invasion and migration of colon cancer cells,and to clarify whether the HMGA1 gene was the target gene of miR-26 a.Methods:The miR-NC(miR-NC group),miR-26 amimics(miR-26 amimics group)and miR-26 ainhibitor(miR-26 ainhibitor group)were transfected into the human colon cancer SW480 cells.RT-PCR and Western blotting methods were used to detect the mRNA and protein expression levels of HMGA1.Luciferase reporter gene detection kit was used to detect the double luciferase activity in SW480 cells and to determine whether HMGA1 was the target gene of miR-26 a.The cell proliferation activities of cells in various groups were detected by CCK8 assay;the number of invasion and migration cells was detected by Transwell chamber method.Results:HMGA1 gene was the target gene of miR-26 a.Compared with miR-NC group,the proliferation activities of colon cancer SW480 cells in miR-26 amimics group at after 24,48 and 72 hafter transfection were significantly decreased(P<0.01),while the proliferation activities of colon cancer SW480 cells in miR-26 ainhibitor group were significantly increased(P<0.01).Compared with miR-NC group,the proliferation activities of colon cancer SW480 cells,the number of invasion and migration cells in miR-26 a mimics group and miR-26 a mimics+pcDNA3.1-HMGA1 group at different time points were significantly decreased(P<0.01);the proliferation activities of colon cancer SW480 cells,the number of invasion and migration cells in miR-NC+pcDNA3.1-HMGA1 group were increased(P<0.01).Compared with miR-26 amimics group,the proliferation activities of colon cancer SW480 cells,the number of invasion and migration cells in miR-26 amimics+pcDNA3.1-HMGA1 group at different time points were increased(P<0.01);compared with miR-NC+pcDNA3.1-HMGA1 group,the proliferation activities of colon cancer SW480 cells,the number of invasion and migration cells in miR-26 mimics+pcDNA3.1-HMGA1 group at different time points were decreased(P<0.01).Conclusion:HMGA1 gene is the target gene of miR-26 a.Up-regulation of the expression of miR-26 acan inhibit the proliferation of colon cancer SW480 cells and downregulation of the expression of miR-26 acan promote the proliferation of colon cancer SW480 cells.更多还原