lncRNA TUG1调控miRNA-145促进结肠癌细胞增殖、迁移和侵袭

目的探讨lncRNA TUG1调控miRNA-145促进结肠癌细胞的增殖、迁移和侵袭。方法选择人结肠癌细胞HCT116，并将细胞分为6组，培养48 h后进行后续实验；采用qRT-PCR法检测各组细胞TUG1与miRNA-145的表达水平；采用MTT检测各组细胞的增殖情况；采用划痕实验和Transwell侵袭实验检测细胞迁移、侵袭能力；采用双荧光素酶报告基因实验验证TUG1与miRNA-145的靶向关系。结果与人正常结肠黏膜上皮细胞株NCM460相比，结肠癌细胞株HCT116中TUG1相对表达水平明显升高，miRNA-145相对表达水平明显降低（P < 0.01）；miRNA-145与TUG1 3'-UTR存在结合位点。与TUG1 siRNA阴性对照组相比，TUG1 siRNA组HCT116细胞中TUG1表达水平明显降低（P < 0.01），HCT116细胞增殖活性降低（P < 0.05），细胞划痕相对宽度明显增加，细胞侵袭数目明显减少（P < 0.05）；与miRNA NC组相比，miRNA-145 mimic组HCT116细胞中miRNA-145表达水平明显升高，HCT116细胞增殖活性降低（P < 0.05），细胞划痕相对宽度明显增加，细胞侵袭数目明显减少（P < 0.01）；与TUG1 siRNA阴性对照组相比，TUG1 siRNA组HCT116细胞中miRNA-145表达水平升高（P < 0.05）；与si-TUG1+miRNA NC组相比，TUG1 siRNA+miRNA-145 inhibitor组HCT116细胞中miRNA-145表达水平降低（P < 0.05），HCT116细胞增殖活性增强，细胞划痕相对宽度减小，细胞侵袭数目增加（P < 0.05）。结论结肠癌HCT116细胞中TUG1表达上调，miRNA-145表达下调，干扰TUG1表达可通过靶向上调miRNA-145的表达，抑制结肠癌HCT116细胞的增殖、迁移和侵袭。

Effect of lncRNA TUG1 regulating miRNA-145 expression on the proliferation,migration and invasion of colon cancer cells

ObjectiveTo explore the effects of lincRNA TUG1 regulating miRNA-145 on the proliferation, migration and invasion of colon cancer cells.MethodsHuman colon cancer cell line HCT116 was divided into 6 groups, and cultured for 48 h.The expression levels of TUG1 and miRNA-145 in each group were detected using qRT-PCR.The proliferation of cells was detected using MTT assay, and the ability of cell migration and invasion were detected using scratch test and Transwell invasion test.The targeting relationship between TUG1 and miRNA-145 was verified by dual-luciferase reporter gene assay.ResultsCompared with human normal colonic mucosal epithelial cell line NCM460, the relative expression level of TUG1 in colon cancer cell line HCT116 significantly increased, while the relative expression level of miRNA-145 significantly decreased(P < 0.01).There were binding sites for miRNA-145 and TUG1 3'-UTR.Compared with the si-con group, the TUG1 expression level and proliferation activity of HCT116 cells in the TUG1 siRNA group significantly decreased, the relative width of cell scratch significantly increased, and the number of invaded cells significantly decreased(P < 0.05).Compared with the miRNA NC group, the miRNA-145 expression level in the miRNA-145 mimic group significantly increased, the proliferation activity of HCT116 cells significantly decreased, the relative width of cell scratch significantly increased, and the number of invaded cells significantly decreased(P < 0.05).Compared with the SI-CON group, the expression level of miRNA-145 in TUG1 siRNA group significantly increased(P < 0.05).Compared with the si-TUG1+miRNA NC group, the expression level of miRNA-145 in the TUG1 siRNA+ mirNA-145 inhibitor group significantly reduced(P < 0.05), the proliferation activity of HCT116 cells significantly enhanced, the relative width of cell scratche significantly reduced, and the number of invaded cell significantly increased(P < 0.05).ConclusionsThe expression level of TUG1 is up-regulated and the expression level of miRNA-145 is down-regulated in colon cancer HCT116 cells.Interfering with the expression of TUG1 can inhibit the proliferation, migration and invasion of colon cancer HCT116 cells by targeting up-regulation of miRNA-145 expression.