Effect of microRNA-451 on proliferation and migration of colorectal carcinoma via inhibition the expression of macrophage migration inhibitory factor

Abstract

Objective To explore effect of microRNA (miRNA, miR)-451 on proliferation and migration of colorectal carcinoma via macrophage migration inhibitory factor (MIF). Methods miR-451 mimics and miR-451 NC were transfected into colorectal carcinoma cell by liposome Lipofectamine™3000. The expression of miR-451 was detected by real-time quantitative polymerase chain reaction (Real-time PCR). Cell viability was measured by methyl thiazol tetrazolium (MTT) assay. Cell clone ability was detected by cell clone formation test. Cell cycle was detected by flow cytometry. Cell migration ability was detected by Transwell method. The expression of MIF protein and mRNA was measured by Western blotting and Real-time PCR. Luciferase reporter analysis was performed. Results The expression of miR-451 in miR-451 mimics group (2.46±0.5) was higher than that in miR-451 NC group (1.00±0.11, t=52.374, P<0.05). Cell viability in miR-451 mimics group (0.37±0.03) was lower than that in miR-451 NC group (0.58±0.06, t=30.672, P<0.05). Cell clone number in miR-451 mimics group (41.56±4.17) was lower than that in miR-451 NC group (132.37±11.29, t=73.927, P<0.05). The G1 period in miR-451 mimics group was longer than that in miR-451 NC group (t=4.093, P<0.05), and the G2 period in miR-451 mimics group was shorter than that in miR-451 NC group (t=6.349, P<0.05). The number of cell invasion in miR-451 mimics group (56.59±5.66) was less than that in the miR-451NC group (184.73±18.47, t=64.992, P<0.05). The fluorescence intensity in miR-451 mimics+ pGL4 MIF 3’untranslated regions (3’UTR)-Wt group was lower than other transfection groups (F=2 875.12, P<0.05). The expression of MIF mRNA in miR-451 mimics (0.35±0.03) group was lower than that in miR-451 NC group (1.00±0.10, t=61.001, P<0.05). The western blot results showed that expression amount of MIF protein in mir-451 mimics group (0.23±0.02) was lower than that in miR-451 NC group (0.92±0.10, t=66.293, P<0.05). Conclusion Over-expression of miR-451 could inhibit colorectal carcinoma cell proliferation and migration by targeting down-regulation expression of MIF. Key words: Carcinoma carcinoma; MicroRNA-451; Macrophage migration inhibitory factor; Proliferation; Migration