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.

中文翻译：

---

小檗碱对人黑色素瘤 A375 细胞的细胞周期调控

我们研究了小檗碱对皮肤黑色素瘤 A375 细胞增殖、凋亡和迁移的影响，以及细胞周期相关的 miRNA 及其靶基因 CDK1、CDK2 和细胞周期蛋白 D1 和 A。小檗碱对皮肤黑色素瘤 A375 细胞增殖、凋亡和迁移的影响。 MTT法评价A375细胞的生长。台盼蓝染色检测细胞凋亡。通过划痕测试评估细胞迁移。通过流式细胞术确定细胞周期阶段。通过 qRT-PCR 测量 miRNA-582-5p 和 miRNA-188-5 及其编码 CDK1、CDK2 和细胞周期蛋白 D1 和 A 的靶基因的 mRNA 的水平。通过蛋白质印迹确定细胞周期相关蛋白（CDK1、CDK2 和细胞周期蛋白 D1 和 A）的表达。小檗碱以时间和剂量依赖性方式抑制 A375 细胞的增殖，并显着且剂量依赖性地增强细胞凋亡。划痕试验显示小檗碱对 A375 细胞迁移有抑制作用。低浓度 (20 和 40 μM) 的小檗碱导致 S 和 G2/M 期的细胞周期停滞，而高浓度的小檗碱 (60 和 80 μM) 处理会导致 G2/M 期的细胞周期停滞。小檗碱浓度的增加导致 miRNA-582-5p 和 miRNA-188-5p 表达增加，以及编码 CDK1、CDK2 和细胞周期蛋白 D1 和 A 的相应靶基因的 mRNA 表达降低。蛋白质印迹还显示降低 CDK1、CDK2 和细胞周期蛋白 D1 和 A 的表达。因此，小檗碱抑制人类黑色素瘤细胞的生长和迁移并促进其凋亡。