Ma Y G, Han Y Z, Zhang Z S, Yu Y, Xu X F, Yuan L
Department of Gastroenterology, the Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
Department of Anatomy, School of Basic Medcine, Henan College of Traditional Chinese Medicine, Zhengzhou 450046, China.
Zhonghua Zhong Liu Za Zhi. 2020 Apr 23;42(4):312-318. doi: 10.3760/cma.j.cn112152-20190924-00622.
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). 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. 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, <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, <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 (<0.01), reversed the effect of miR-451 suppressed proliferation and migration of SW480 cells. MiR-451 may regulate proliferation and migration of human colorectal cancer cells by targeting MIF.
通过靶向巨噬细胞移动抑制因子（MIF）来研究miR-451对人结肠癌细胞SW480增殖和迁移的影响及机制。采用基因芯片分析筛选SW480细胞中差异表达的微小RNA和信使核糖核酸。运用实时定量聚合酶链反应（RT-qPCR）检测转染前后SW480细胞中miR-451和MIF的表达。分别采用细胞克隆形成实验和Transwell实验检测SW480细胞的增殖和侵袭能力。运用细胞划痕实验检测SW480细胞的迁移能力。利用TargetScan数据库分析miR-451与MIF之间的相关性。采用双荧光素酶报告基因检测miR-451与MIF的相互作用。运用MTT实验检测SW480细胞的活力。与正常人结肠黏膜细胞FHC（1.00）相比，SW480细胞中miR-451的表达下调（0.36±0.18，<0.001）。敲低miR-451可促进SW480细胞的增殖和迁移。与FHC细胞相比，SW480细胞中MIF表达上调（2.28±0.45，<0.001）。下调MIF可抑制SW480细胞的增殖、侵袭和迁移。miR-451特异性结合MIF的3'非翻译区并调节MIF的表达。过表达miR-451可降低SW480细胞的活力，而过表达MIF则可增加其活力。过表达MIF促进SW480细胞的增殖和迁移（<0.01），逆转了miR-451抑制SW480细胞增殖和迁移的作用。miR-451可能通过靶向MIF调节人结肠癌细胞的增殖和迁移。
