Cell Cycle Regulation by Berberine in Human Melanoma A375 Cells
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Bulletin of Experimental Biology and Medicine, Vol. 169, No. 4, August, 2020 ONCOLOGY
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Cell Cycle Regulation by Berberine in Human Melanoma A375 Cells
Min Ren1, Lihui Yang1, Dongxia Li1, Ling Yang2, Yila Su2, and Xiulan Su2 Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 169, No. 4, pp. 477-483, April, 2020 Original article submitted May 27, 2019 We studied the effects of berberine on the proliferation, apoptosis, and migration of skin melanoma A375 cells, as well as cell cycle-related miRNAs and their target genes, CDK1, CDK2, and cyclins D1 and A. The inhibitory effect of berberine on the growth of A375 cells was evaluated by MTT assay. Cell apoptosis was detected by trypan blue staining. Cell migration was assessed by the scratch test. Cell cycle phases were determined by flow cytometry. The levels of miRNA-582-5p and miRNA-188-5, and mRNA of their target genes encoding CDK1, CDK2, and cyclins D1 and A were measured by qRT-PCR. The expression of cell cycle-related proteins (CDK1, CDK2, and cyclins D1 and A) was determined by Western blotting. Berberine inhibited the proliferation of A375 cells in a time- and dose-dependent manner and significantly and dose-dependently enhanced cell apoptosis. Scratch assay showed an inhibitory effect of berberine on migration of A375 cells. Berberine in low concentrations (20 and 40 μM) caused cell cycle arrest in the S and G2/M phases, while treatment with high concentrations of berberine (60 and 80 μM) arrested cell-cycle in the G2/M phase. The increase in berberine concentration led to an increase in miRNA-582-5p and miRNA-188-5p expression and a decrease in the expression of mRNA for the corresponding target genes encoding CDK1, CDK2, and cyclins D1 and A. Western blotting also revealed reduced expression of CDK1, CDK2, and cyclins D1 and A. Thus, berberine suppressed the growth and migration of human melanoma cells and promoted their apoptosis. Berberine can increase the expression of cell cycle-related miRNAs and cause degradation of the corresponding target genes, thereby blocking the cell cycle progression and inhibiting the melanoma A375 cells. Key Words: berberine; melanoma; A375 cells; miRNA; cell cycle-related proteins Melanoma is the most aggressive skin tumor with high malignancy, metastasis, and mortality. It arises from melanocytes, mostly present in the skin and also found in the pia mater and eye choroid. At the early stages, melanoma can be treated by surgery, but advanced or metastatic melanoma is insensitive to radio-, chemo-, or biotherapy and has poor prognosis [6]. Moreover, toxic side effects of therapy determine poor patient compliance and seriously impair the quality of life. Therefore, Chinese herbal medicine with minimum Department of Dermatology, Affiliated Hospital of Inner Mongolia Medical University, Huhhot, Inner Mongolia, P.R. China; 2Clinical Medical Research Center, Inner Mongolia Medical University, Huhhot, Inner Mongolia, P.R. China. Address for correspondence: [email protected]. Dongxia Li 1
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