A study on mechanism underlying the inhibition of berberine on proliferation of human melanoma A375 cells / 中华皮肤科杂志

ABSTRACT

Objective To evaluate the effect of berberine on the expression of cell cycle-related miRNA and its target gene in human melanoma A375 cells.Methods In vitro cultured A375 cells were classified into several groups to be treated with berberine at different concentrations of 0 (blank control group),20,40,60,80,and 100 μmol/L,respectively,for 48 hours.qRT-PCR was performed to determine the mRNA expression of miRNA-582-5p and its target gene cyclin-dependent kinase 1 (CDK1),miRNA-188-5p and its target gene CDK2,Cyclin D1 and Cyclin A.Western blot analysis was conducted to measure the protein expression of cell cycle-related proteins CDK1,CDK2,Cyclin D1 and Cyclin A.Results qRT-PCR showed that compared with the blank control group,the 20 and 40 μmol/L berberine groups had a similar expression of miRNA-582-5p (both P ＞ 0.05),but the 60 and 80 μmol/L berberine groups had a significantly up-regulated expression of miRNA-582-5p (both P ＜ 0.05).Compared with the blank control group,all the 5 berberine groups had a significantly increased expression of miRNA-188-5p (F =22.600,P =0.002).However,the mRNA expression of CDK2,CDK1,Cyclin A and Cyclin D1 gradually decreased along with the increase of berberine concentrations (F =51.976,248.510,626.671 and 312.740,respectively,all P ＜ 0.001).Western blot analysis revealed that berberine decreased the protein expression of CDK1,CDK2,Cyclin D1 and Cyclin A (F =138.124,110.966,278.772 and 140.167,all P ＜ 0.001).Conclusion Berberine can decrease the expression of cell cycle-related proteins CDK1,CDK2,Cyclin D1 and Cyclin A,likely by decreasing their mRNA expression and increasing the expression of miRNA-582-5p and miRNA-188-5p,and then block the cell cycle progression of A375 cells and inhibit the growth of tumor.