Mechanism Study on Proliferation and Migration of Keloid Fibroblasts Regulated by Psoralen Isoflavones Via miR-497-5p
Objective To investigate the mechanism of psoralen isoflavones inhibiting the proliferation and migration of keloid fibroblasts by regulating miR-497-5p.Methods Keloid fibroblasts were cultured in vitro and treated with 0,9,18,36 μg/ml psoralen isoflavones,which were classified as Con group,low dose group,medium dose group and high dose group.The cells were transfected with miR-NC and miR-497-5p by liposome method,and were denoted as miR-NC group and miR-497-5p group.After transfection with anti-miR-NC and anti-miR-497-5p by liposome method,the cells were treated with 36 μg/ml psoralen isoflavones,which were denoted as high-dose group+anti-miR-NC group and high-dose group+anti-miR-497-5p group.Cell proliferation was detected by CCK8 and cloning assay.Cell migration was detected by wound healing test.The expression levels of Cyclin D1,MMP2 and MMP9 were detected by Western blot assay.The expression level of miR-497-5p was detected by qRT-PCR.Results Psoralen isoflavones decreased the cell viability and migration rate of keloid fibroblasts(P＜0.05),decreased the number of cloned cells(P＜0.05),and decreased the protein expression levels of Cyclin D1,MMP2,and MMP9(P＜0.05),increase the expression level of miR-497-5p.Compared with the miR-NC group,the cell viability and migration rate of the miR-497-5p group were reduced(P＜0.05),the number of cloned cells was reduced(P＜0.05),and the protein expression levels of Cyclin D1,MMP2,and MMP9 were reduced(P＜0.05).Inhibition of miR-497-5p can reverse the effect of psoralen isoflavones on the proliferation and migration of keloid fibroblasts.Conclusion Psoralen isoflavones may inhibit the proliferation and migration of keloid fibroblasts by increasing miR-497-5p.
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