MicroRNA-182 regulates colon cancer cell proliferation and migration by targeting special AT-rich sequence-binding protein 2 gene

Abstract

Objective To observe the effect of microRNA (miRNA, miR)-182 on the proliferation and migration of colorectal cancer cells by targeting special AT-rich sequence-binding protein 2 (SATB2) gene, and to explore its effect on SATB2/icotinamide adenine dinucleotide phosphate oxidase 4 (nox4) pathway. Methods HT-29 cells in logarithmic phase were transfected with Si-miR-182 and Control-si resectively, by liposome transfection, serving as Si-miR-182 group and N-miR-182 group, respectively. The expression of miR-182 gene, the cell proliferation and migration, the mRNA and protein expression of SATB2, nox4, E-cadherin and vimentin were examined. Results The relative expression of miR-182 gene in Si-miR-182 group (0.35±0.05) was lower than that in control group (1.24±0.26) and N-miR-182 group (1.20±0.25) (t=7.517, 7.455, P<0.05). The A values of methyl thiazol tetrazolium (MTT) test in Si-miR-182 group were lower than those in control group and N-miR-182 group after 24, 48 and 72 h of culture (24 h: t=2.667, 2.664; 48 h: t=4.559, 4.524; 72 h: t=7.257, 6.981; P<0.05). Compared with 24-h culture, the A value of MTT test in 48 and 72 hours in three groups were increased (48 h: t=5.507, 5.092, 3.741; 72 h: t=11.330, 10.637, 9.229; P<0.05), and the A value of MTT test in 72 hours were higher than those in 48 hours(t=7.411, 6.941, 5.214, P<0.05). The cell migration rate of Si-miR-182 group [(53.90±3.19)%] was lower than that of the control group [(81.66±5.92)%] and N-miR-182 group [(80.35±5.40)%] (t=9.231, 9.430, P<0.05). The relative expressions of SATB2, E-cadherin mRNA and protein in Si-miR-182 group were higher than those in control group and N-miR-182 group (SATB2: t=10.930, 11.158; E-Cadherin: t=9.288, 9.369; P<0.05), while the relative expressions of nox4, vimentin mRNA and protein in Si-miR-182 group were lower than those in control group and N-miR-182 group (nox4: t=8.955, 7.590; Vimentin: t=6.543, 6.644; P<0.05). Conclusion Silencing of miR-182 gene can significantly inhibit the proliferation and migration of colorectal cancer cells, possibly by activating SATB2, E-Cadherin expression, inhibiting the expression of nox4, Vimentin, and participating in the SATB2/nox4 pathway. Key words: MicroRNA-182; Special AT-rich sequence-binding protein 2; Colorectal cancer; Proliferation; Migration