Expression of miR-489 in keloid tissues and the effect of its overexpression on proliferation,migration and fibrosis of keloid fibroblasts
Objective To observe the expression changes of miR-489 in the keloid tissues and fibroblasts,and to ex-plore the effect of miR-489 overexpression on the proliferation,migration and fibrosis of keloid fibroblasts.Methods Six cases of keloid tissues and six cases of normal skin tissues were collected.Primary keloid fibroblasts and normal fibroblasts were isolated and subcultured.The relative expression of miR-489 in tissues and cells was detected by real-time fluores-cence quantitative PCR.Keloid fibroblasts were divided into the control group and overexpression group,which were trans-fected with miR-NC and miR-489-mimic,respectively.After 48 h of transfection,CCK-8 method was used to detect the cell proliferation ability after continuous culture for 5 days(expressed as OD450),and Transwell chamber experiment was used to detect the cell migration ability(expressed as the number of migration cells).The relative expression levels of type I collagen(COL1),type III collagen(COL3)antibody,α-smooth muscle actin(α-SMA)and Smad3 protein were detect-ed by Western blotting.Results The relative expression levels of miR-489 in the keloid tissues and normal skin tissues were 0.50±0.06 and 1.00±0.00,respectively,with statistically significant difference(both P<0.05).The relative ex-pression levels of miR-489 in keloid fibroblasts and normal fibroblasts were 0.25±0.18 and 1.00±0.00,respectively,with statistically significant difference(both P<0.05).The cell proliferation ability of the overexpression group was lower than that of the control group on the 3rd to 5th days of culture(P<0.05).There was no significant difference in cell prolif-eration ability between the two groups on the 1st and 2nd days of culture(P>0.05).The numbers of migration cells in the overexpression group and the control group were 43.00±4.00 and 129.67±9.71,respectively(P<0.05).The relative expression levels of COL1,COL3,α-SMA and Smad3 proteins in the overexpression group were lower than those in the control group(all P<0.05).Conclusions The expression of miR-489 in keloid tissues and fibroblasts decreased.Over-expression of miR-489 may inhibit the proliferation,migration and fibrosis of keloid fibroblasts by regulating the expression of target gene Smad3 protein.
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