Grape seed proanthocyanidins inhibit melanoma cell invasiveness by reduction of PGE2 synthesis and reversal of epithelial-to-mesenchymal transition.
Grape seed proanthocyanidins inhibit melanoma cell invasiveness by reduction of PGE2 synthesis and reversal of epithelial-to-mesenchymal transition.
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DOI：
10.1371/journal.pone.0021539
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Katiyar SK
中科院分区：
文献类型：
--
作者：
Vaid M;Singh T;Katiyar SK
黑色素瘤是皮肤病的主要死亡原因，在很大程度上是因为它容易转移。我们研究了葡萄籽原花青素(GSPs)对黑色素瘤癌细胞迁移的影响以及这些作用的分子机制。通过体外细胞侵袭实验，我们观察到GSPs对A375和Hs294t细胞的侵袭和迁移有浓度依赖性的抑制作用，这与环氧合酶(COX)-2的表达水平和前列腺素(PG)E2的产生减少有关。用环氧合酶-2抑制剂塞来昔布处理细胞，或用环氧合酶-2小干扰RNA瞬时转染黑色素瘤细胞，也能抑制黑色素瘤细胞的迁移。用COX-2的诱导剂12-O-十四酰佛波醇-13-乙酸酯处理细胞，可促进丝裂原活化蛋白激酶家族蛋白ERK1/2的磷酸化，进而促进细胞迁移，而GSPs和塞来昔布均显著抑制12-O-十四酰佛波醇-13-乙酸酯促进的细胞迁移和ERK1/2的磷酸化。MEK抑制剂UO126处理细胞也抑制黑色素瘤细胞迁移。此外，GSP抑制了黑色素瘤细胞中COX-2的上游调节因子--NF-κB/p65的激活，并且用NF-κB的抑制剂咖啡酸苯乙酯处理细胞也抑制了细胞的迁移。此外，GSP抑制黑色素瘤细胞的迁移与逆转上皮-间充质转化过程有关，这导致黑色素瘤细胞中上皮生物标志物(E-钙粘蛋白和细胞角蛋白)水平增加，而间质生物标志物(波形蛋白、纤维连接蛋白和N-钙粘附素)丢失。综上所述，这些结果表明GSPs有能力通过靶向COX-2的内源性表达和逆转上皮向间充质转化的过程来抑制黑色素瘤细胞的侵袭/迁移。
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 grape seed proanthocyanidins (GSPs) on melanoma cancer cell migration and the molecular mechanisms underlying these effects using highly metastasis-specific human melanoma cell lines, A375 and Hs294t. Using in vitro cell invasion assays, we observed that treatment of A375 and Hs294t cells with GSPs resulted in a concentration-dependent inhibition of invasion or cell migration of these cells, which was associated with a reduction in the levels of cyclooxygenase (COX)-2 expression and prostaglandin (PG) E2 production. Treatment of cells with celecoxib, a COX-2 inhibitor, or transient transfection of melanoma cells with COX-2 small interfering RNA, also inhibited melanoma cell migration. Treatment of cells with 12-O-tetradecanoylphorbol-13-acetate, an inducer of COX-2, enhanced the phosphorylation of ERK1/2, a protein of mitogen-activated protein kinase family, and subsequently cell migration whereas both GSPs and celecoxib significantly inhibited 12-O-tetradecanoylphorbol-13-acetate -promoted cell migration as well as phosphorylation of ERK1/2. Treatment of cells with UO126, an inhibitor of MEK, also inhibited the migration of melanoma cells. Further, GSPs inhibited the activation of NF-κB/p65, an upstream regulator of COX-2, in melanoma cells, and treatment of cells with caffeic acid phenethyl ester, an inhibitor of NF-κB, also inhibited cell migration. Additionally, inhibition of melanoma cell migration by GSPs was associated with reversal of epithelial-mesenchymal transition process, which resulted in an increase in the levels of epithelial biomarkers (E-cadherin and cytokeratins) while loss of mesenchymal biomarkers (vimentin, fibronectin and N-cadherin) in melanoma cells. Together, these results indicate that GSPs have the ability to inhibit melanoma cell invasion/migration by targeting the endogenous expression of COX-2 and reversing the process of epithelial-to-mesenchymal transition.