Effect of inhibiting the miR-31 expression on the proliferation and apoptosis of human keratinocyte cells

ObjectiveTo investigate the effects of inhibiting the miR-31 expression on the proliferation and apoptosis of human keratinocyte cells, and its mechanism.

MethodsThe HaCaT cells were cultured, and transfected using transient transfection method.These cells were divided into the antisense oligonueleotide(ASO) group(treatment with miR-31 ASO), control ASO group(treatment with control ASO) and blank control group(treatment with blank plasmid).The cell proliferation, apoptosis, cell cycle and Rho related domain BTB protein 1(RhoBTB1) expression in three groups were detected using MTT assay, Annexin V-FITC/PI double staining, PI single staining and Western blotting, respectively.

ResultsThe expression level of miR-31 in ASO group were significantly lower than that in ASO control group and blank control group(P < 0.01).The absorbance values in ASO group at 24 h, 48 h, 72 h and 96 h were significantry lower than that in ASO control group and blank control group(P < 0.05 to P < 0.01), and the absorbance values in three groups gradually increased as time went on(P < 0.01).The apoptotic rate, proportion of G₀/G₁ phase cells and expression level of RhoBTB1 protein in ASO group were significantly higher than those in ASO control group and blank control group(P < 0.01), while the proportion of S/G₂ phase cells in ASO group were significantly lower than those in ASO control group and blank control group(P < 0.01).

ConclusionsmiR-31 may be involved in the proliferation and apoptosis of human keratinocyte cell lines HaCaT cells, which may achieve this biological process by regulating the expression of RhoBTB1 protein.