上调miR-433表达抑制结直肠癌细胞系增殖

Up-regulation of miR-433 expression inhibits proliferation of colorectal cancer cell line in vitro

目的研究miR-433在结直肠癌中的表达,并初步探讨其功能。方法选择不同分期结直肠癌癌组织(CRC)、癌旁组织与正常结直肠组织,用实时定量PCR检测miR-433表达并比较;体外培养结直肠癌细胞系HT-29、HCT-116和SW480以及正常结直肠细胞系NCM460,用实时定量PCR检测miR-433表达。随后将miR-433模拟物转染SW480细胞,CCK-8法检测细胞增殖;划痕实验检测细胞迁移;Transwell小室法检测细胞侵袭;流式细胞测量术检测细胞周期。结果与癌旁组织相比,miR-433在结直肠癌组织中表达显著降低(P<0.05)。在结肠癌患者血清中miR-433水平也显著低于健康人群(P<0.05)。此外,从TNM分期Ⅰ期~Ⅳ期,结直肠癌组织内miR-433表达逐渐降低(P<0.05)。与NCM460相比,HT-29、HCT-116和SW480中,miR-433表达下调(P<0.05)。SW480转染miR-433模拟物后,与对照miRNA组相比,在SW480细胞增殖水平明显降低(P<0.05),G2和M期细胞比例增多。上调miR-433后,细胞侵袭和迁移明显减弱(P<0.05)。结论miR-433在结直肠癌中表达降低。在SW480细胞中上调其表达后,可抑制细胞系增殖、侵袭与迁移。

Objective To study the expression and function of miR-433 in the development of colorectal cancer.Methods RT-qPCR was used to detect the expression of miR-433 in clinical specimens and serum of patients with colorectal cancer,and compared with those from adjacent tissues or at different stages of tumore development and from normal persons.The colorectal cancer cell lines HT-29,HCT-116,SW480 and normal colon cell line NCM460 were cultured,and the expression of miR-433 was measured by real-time quantitative PCR.Subsequently,miR-433 mimic was transfected into SW480 cells,CCK-8 was used to detect cell proliferation,flow cytometry was used to detect cell cycle changes,scratch test and Transwell test were used to detect cell migration and invasionrespectively.Results Compared with the adjacent tissues,the expression of miR-433 in colorectal cancer tissues was significantly decreased(P<0.05).The serum level of miR-433 in patients with colon cancer was also significantly lower than that in normal persons(P<0.05).In addition,from stageⅠto stageⅣof TNM,the expression of miR-433 in colorectal cancer tissues gradually and significantly decreased.Compared with normal human colon epithelial cell line NCM460,the expression of miR-433 was down-regulated in colorectal cancer cell lines HT-29,HCT-116 and SW480(P<0.05).Compared with the control group,the proliferation of SW480 cells was significantly inhibited(P<0.05)and the proportion of G2 and M phase cells increased after SW480 transfection.The invasion and migration of cells decreased significantly after up-regulation of miR-433(P<0.05).Conclusions The expression of miR-433 is decreased in colorectal cancer.Up-regulation of its expression in SW480 cells inhibits cell proliferation,invasion and migration.