miR-451靶向巨噬细胞移动抑制因子对结直肠癌细胞增殖迁移的影响及其作用机制

MiR-451 regulates proliferation and migration of colorectal cells by targeting MIF

目的:探讨miR-451通过靶向巨噬细胞移动抑制因子(MIF)对人结直肠癌细胞SW480增殖和迁移的影响及其作用机制。方法:微阵列分析筛选SW480细胞中差异表达的微小RNA和信使RNA,实时定量PCR法检测miR-451和MIF在SW480细胞中的表达以及转染miR-451、MIF后miRNA-451和MIF的表达,细胞克隆形成实验检测miR-451和MIF的表达对SW480细胞增殖能力的影响,Transwell实验检测miR-451和MIF的表达对SW480细胞侵袭能力的影响,细胞划痕实验检测miR-451和MIF的表达对SW480细胞迁移能力的影响,TargetScan数据库分析miR-451与MIF的相关性,双荧光素酶报告基因检测miR-451与MIF的相互作用,四甲基偶氮唑蓝法检测MIF过表达对SW480细胞活力的影响。结果:与人正常结直肠黏膜细胞FHC(1.00)比较,SW480细胞中miR-451的表达(0.36±0.18)下调( P<0.001)。miR-451过表达抑制SW480细胞的增殖和迁移,miR-451低表达促进SW480细胞的增殖和迁移。与人正常结直肠黏膜细胞FHC(1.00)比较,SW480细胞中MIF的表达(2.28±0.45)上调( P<0.001);MIF低表达抑制SW480细胞的增殖、侵袭和迁移。miR-451与MIF 3′UTR特异性结合,调控MIF的表达活性。miR-451过表达降低SW480细胞的活力,MIF过表达使SW480细胞的活力增加。MIF过表达能促进SW480细胞的增殖和迁移( P<0.01),MIF过表达逆转miR-451过表达对SW48细胞增殖和迁移的作用。 结论:miR-451通过靶向MIF可能调控人结直肠癌细胞的增殖和迁移。

Objective To investigate the effect and mechanism of miR-451 on the proliferation and migration of human colorectal cancer cell SW480 by targeting macrophage migration inhibitory factor(MIF).Methods Microarray analysis was used to screen differentially expressed microRNAs and messenger RNA in SW480 cells.Real-time quantitative PCR(RT-qPCR)was used to detect the expressions of miR-451 and MIF in SW480 cells before and after transfection.Cell clone formation assay and Transwell assay were used to detect the proliferation and invasion of SW480 cells,respectively.Cell scratch assay was used to detect the migration ability of SW480 cells.The TargetScan database was used to analyze the correlation between miR-451 and MIF.Dual luciferase reporter gene was used to detect the interaction of miR-451 and MIF.MTT assay was used to detect the viability of SW480 cells.Results Compared with human normal colorectal mucosal cell FHC(1.00),the expression of miR-451 was down-regulated in SW480 cells(0.36±0.18,P<0.001).Knockdown of miR-451 promoted proliferation,and migration of SW480 cells.Compared with that in FHC cells,MIF expression was up-regulated in SW480 cells(2.28±0.45,P<0.001).MIF down-regulation inhibited SW480 cell proliferation,invasion and migration.MiR-451 specifically bind to the MIF 3′UTR and regulated the expression of MIF.Overexpression of miR-451 reduced while overexpression of MIF increased the viability of SW480 cells.Overexpression of MIF promoted the proliferation and migration of SW480 cells(P<0.01),reversed the effect of miR-451 suppressed proliferation and migration of SW480 cells.Conclusion MiR-451 may regulate proliferation and migration of human colorectal cancer cells by targeting MIF.