上调miR-200a通过抑制MACC1影响结直肠癌细胞生物学行为的研究

Up-regulation of miR-200a Affecting the Biological Behavior of Colorectal Cancer Cells by Inhibiting MACC1

目的探讨上调miR-200a表达是否可以通过抑制MACC1影响结直肠癌LoVo细胞的生物学行为。方法结直肠癌细胞LoVo分组处理为mimics组(转染miR-200a mimics)、blank组(转染空脂质体)和NC组(转染negative control)。转染成功后,CCK-8法、黏附实验和Transwell实验分别检测癌细胞增殖、同质黏附能力和侵袭能力的变化情况;MACC1蛋白表达用Western blot检测;生物信息学预测miR-200a与MACC1靶向情况。结果CCK-8法显示,mimics组细胞的OD值为(1.27±0.07),比blank组和NC组均显著低,差异有统计学意义(F=27.780,P<0.001);mimics组同质性肿瘤细胞黏附数量为(1241.33±76.42),细胞黏附能力比blank组和NC组均显著增加,差异有统计学意义(F=13.130,P=0.006);mimics组的侵袭穿膜细胞数为(54.20±6.83),比blank组和NC组均显著减低,差异有统计学意义(F=13.130,P=0.006);Western Blot均显示mimics组的MACC1蛋白表达显著低于其他两组,差异有统计学意义(F=32.084,P=0.001)。数据库显示MACCI是miR-200a的潜在靶基因。结论上调miR-200a表达可能通过靶向抑制MACC1表达阻遏人结直肠癌细胞的增殖和侵袭能力,增强细胞黏附力。

Objective To investigate whether up-regulation of miR-200a expression can affect the biological behavior of colorectal cancer LoVo cells by inhibiting MACC1.Methods Colorectal cancer cells LoVo were divided into mimics group(transfected with miR-200a mimics),blank group(transfected with empty liposomes)and NC group(transfected with negative control).After successful transfection,CCK-8 method,adhesion experiment and Transwell experiment were used to detect the changes of cancer cell proliferation,homogenous adhesion ability and invasion ability;MACC1 protein expression was detected by Western blot;bioinformatics predicted miR-200a and MACC1 targets situation.Results The CCK-8 method showed that the OD value of the mimics group was(1.27±0.07),which was significantly lower than that of the blank group and the NC group,the difference was statistically significant(F=27.780,P<0.001);the number of homogeneous tumor cells in the mimics group was(1241.33±76.42),the cell adhesion ability was significantly increased,compared with the blank group and the NC group,the difference was statistically significant(F=13.130,P=0.006);the number of invasion and penetrating cells in the mimics group was(54.20±6.83),which was significantly lower than that of the blank group and the NC group,the difference was statistically significant(F=13.130,P=0.006);Western Blot showed that the MACC1 protein expression in the mimics group was significantly lower than the other two groups,the difference was statistically significant(F=32.084,P=0.001).The database shows that MACCI was a potential target gene of miR-200a.Conclusion Up-regulating the expression of miR-200a may inhibit the proliferation and invasion of human colorectal cancer cells by targeting the expression of MACC1,and enhance cell adhesion.