小檗碱通过影响 FA K、uPA 和 NF-κB 信号通路抑制人黑色素瘤 A375.S2 细胞迁移和侵袭，并抑制 PLX4032 耐药 A375.S2 细胞迁移。

Berberine Inhibits Human Melanoma A375.S2 Cell Migration and Invasion via Affecting the FAK, uPA, and NF-κB Signaling Pathways and Inhibits PLX4032 Resistant A375.S2 Cell Migration In Vitro.

机构信息

Department of Biological Science and Technology, China Medical University, Taichung 40402, Taiwan.

Department of Medical Laboratory Science and Biotechnology, College of Medicine and Life Science, Chung Hwa University of Medical Technology, Tainan 71703, Taiwan.

出版信息

Molecules. 2018 Aug 13;23(8):2019. doi: 10.3390/molecules23082019.

Many studies have demonstrated that berberine inhibited the cell migration and invasion in human cancer cell lines. However, the exact molecular mechanism of berberine inhibiting the cell migration and invasion of human melanoma A375.S2 and A375.S2/PLX (PLX4032 induced resistant A375.S2) skin cancer cells remains unknown. In this study, we investigated the anti-metastasis mechanisms of berberine in human melanoma cancer A375.S2 cells and A375.S2/PLX resistant cells in vitro. Berberine at low concentrations (0, 1, 1.5 and 2 μM) induced cell morphological changes and reduced the viable cell number and inhibited the mobility, migration, and invasion of A375.S2 cells that were assayed by wound healing and transwell filter. The gelatin zymography assay showed that berberine slightly inhibited MMP-9 activity in A375.S2 cells. Results from western blotting indicated that berberine inhibited the expression of MMP-1, MMP-13, E-cadherin, N-cadherin, RhoA, ROCK1, SOS-1, GRB2, Ras, p-ERK1/2, p-c-Jun, p-FAK, p-AKT, NF-κB, and uPA after 24 h of treatment, but increased the PKC and PI3K in A375.S2 cells. PLX4032 is an inhibitor of the BRAFV600E mutation and used for the treatment of cancer cells harboring activated BRAF mutations. Berberine decrease cell number and inhibited the cell mobility in the resistant A375.S2 (A375.S2/PLX, PLX4032 generated resistant A375.S2 cells). Based on these observations, we suggest that the potential of berberine as an anti-metastatic agent in melanoma that deserves to be investigated in more detail, including in vivo studies in future.

许多研究表明小檗碱抑制人癌细胞系的细胞迁移和侵袭。然而，小檗碱抑制人黑色素瘤 A375.S2 和 A375.S2/PLX（PLX4032 诱导的 A375.S2 耐药细胞）皮肤癌细胞迁移和侵袭的确切分子机制尚不清楚。在这项研究中，我们研究了小檗碱在体外人黑色素瘤 A375.S2 细胞和 A375.S2/PLX 耐药细胞中的抗转移机制。低浓度（0、1、1.5 和 2 μM）的小檗碱诱导细胞形态发生变化，减少活细胞数量，并抑制 A375.S2 细胞的迁移、迁移和侵袭，通过划痕愈合和 Transwell 滤器进行测定。明胶酶谱分析显示小檗碱轻度抑制 A375.S2 细胞中的 MMP-9 活性。Western blot 结果表明，小檗碱抑制 MMP-1、MMP-13、E-钙黏蛋白、N-钙黏蛋白、RhoA、ROCK1、SOS-1、GRB2、Ras、p-ERK1/2、p-c-Jun、p-FAK、p-AKT、NF-κB 和 uPA 的表达，作用 24 h 后 A375.S2 细胞，但增加 PKC 和 PI3K。PLX4032 是 BRAFV600E 突变的抑制剂，用于治疗携带激活 BRAF 突变的癌细胞。小檗碱减少耐药 A375.S2（A375.S2/PLX，PLX4032 生成耐药 A375.S2 细胞）中的细胞数量并抑制细胞迁移。基于这些观察结果，我们认为小檗碱作为一种抗黑色素瘤转移剂具有潜力，值得更详细地研究，包括未来的体内研究。