Department of Dermatology, University of Alabama at Birmingham, 35294, USA.
Carcinogenesis. 2011 Jan;32(1):86-92. doi: 10.1093/carcin/bgq215. Epub 2010 Oct 25.
Melanoma is the leading cause of death from skin disease due, in large part, to its propensity to metastasize. We have examined the effect of berberine, an isoquinoline alkaloid, on human melanoma cancer cell migration and the molecular mechanisms underlying these effects using melanoma cell lines, A375 and Hs294. Using an in vitro cell migration assay, we show that over expression of cyclooxygenase (COX)-2, its metabolite prostaglandin E₂ (PGE₂) and PGE₂ receptors promote the migration of cells. We found that treatment of A375 and Hs294 cells with berberine resulted in concentration-dependent inhibition of migration of these cells, which was associated with a reduction in the levels of COX-2, PGE₂ and PGE₂ receptors (EP2 and EP4). Treatment of cells with celecoxib, a COX-2 inhibitor, or transient transfection of cells with COX-2 small interfering RNA, also inhibited cell migration. Treatment of the cells with 12-O-tetradecanoylphorbol-13-acetate (TPA), an inducer of COX-2 or PGE₂, enhanced cell migration, whereas berberine inhibited TPA- or PGE₂-promoted cell migration. Berberine reduced the basal levels as well as PGE₂-stimulated expression levels of EP2 and EP4. Treatment of the cells with the EP4 agonist stimulated cell migration and berberine blocked EP4 agonist-induced cell migration activity. Moreover, berberine inhibited the activation of nuclear factor-kappa B (NF-κB), an upstream regulator of COX-2, in A375 cells, and treatment of cells with caffeic acid phenethyl ester, an inhibitor of NF-κB, inhibited cell migration. Together, these results indicate for the first time that berberine inhibits melanoma cell migration, an essential step in invasion and metastasis, by inhibition of COX-2, PGE₂ and PGE₂ receptors.
黑色素瘤是皮肤疾病导致死亡的主要原因，部分原因是其易于转移。我们研究了小檗碱（一种异喹啉生物碱）对人类黑色素瘤癌细胞迁移的影响及其潜在机制，使用了黑色素瘤细胞系 A375 和 Hs294。通过体外细胞迁移实验，我们发现环氧合酶（COX）-2 的过表达、其代谢产物前列腺素 E₂（PGE₂）和 PGE₂ 受体促进了细胞迁移。我们发现，小檗碱处理 A375 和 Hs294 细胞会导致细胞迁移的浓度依赖性抑制，这与 COX-2、PGE₂ 和 PGE₂ 受体（EP2 和 EP4）水平的降低有关。COX-2 抑制剂塞来昔布或 COX-2 小干扰 RNA 瞬时转染处理细胞也抑制细胞迁移。用 12-O-十四烷酰佛波醇-13-乙酸酯（TPA）处理细胞（COX-2 或 PGE₂的诱导剂）可增强细胞迁移，而小檗碱抑制 TPA 或 PGE₂ 促进的细胞迁移。小檗碱降低了 EP2 和 EP4 的基础水平和 PGE₂ 刺激的表达水平。用 EP4 激动剂处理细胞可刺激细胞迁移，小檗碱可阻断 EP4 激动剂诱导的细胞迁移活性。此外，小檗碱抑制了 A375 细胞中 COX-2 的上游调节因子核因子-κB（NF-κB）的激活，NF-κB 的抑制剂咖啡酸苯乙酯抑制了细胞迁移。综上所述，这些结果首次表明，小檗碱通过抑制 COX-2、PGE₂ 和 PGE₂ 受体来抑制黑色素瘤细胞迁移，这是侵袭和转移的重要步骤。
