Liu Jian-Feng, Crépin Michel, Liu Jian-Miao, Barritault Denis, Ledoux Dominique
Laboratoire de Recherche sur la Croissance Cellulaire, la Réparation et la Régénération Tissulaires, CNRS UPRES-A 7053, Université Paris XII, Avenue du Général de Gaulle, 94000 Créteil, France.
Biochem Biophys Res Commun. 2002 May 17;293(4):1174-82. doi: 10.1016/S0006-291X(02)00350-9.
Matrix metalloproteinases (MMPs) play an important role in cancer metastasis. Here, we investigated the effect of fibroblast growth factor-2 (FGF-2) and 12-O-tetradecanoylphorbol-13-acetate (TPA) on the secretion of type IV collagenases (MMP-2, MMP-9) in breast cancer MCF-7 cells. As shown by gelatin zymography, both FGF-2 and TPA stimulated the secretion of MMP-9 in MCF-7 cells while they did not change the level of MMP-2 secretion. Signaling cascade studies indicated that both FGF-2 and TPA induced Ras activation, c-Raf phosphorylation, mitogen-activated protein kinase/ERK kinase (MEK(1/2)) phosphorylation, and extracellular signal-regulated kinase (ERK(1/2)) phosphorylation. The FGF-2- and TPA-induced MMP-9 secretion was significantly inhibited by transient transfection of MCF-7 cells with dominant negative Ras (Ras-N17) and by treatment with MEK(1/2) inhibitor PD98059. A pan-protein kinase C (PKC) inhibitor, GF109203X, was found to totally abolish the FGF-2- and TPA-induced MMP-9 secretion and ERK(1/2) phosphorylation. Use of isoform-specific PKC inhibitors such as Rotllerin and Gö6976 suggested, moreover, that the PKC-delta isoform is a likely component of FGF-2 and TPA trophic signaling. These results demonstrated that FGF-2 and TPA induce MMP-9 secretion in MCF-7 cells mainly through PKC-dependent activation of the Ras/ERK(1/2) signaling pathway.
基质金属蛋白酶（MMPs）在癌症转移中起重要作用。在此，我们研究了成纤维细胞生长因子-2（FGF-2）和12-O-十四烷酰佛波醇-13-乙酸酯（TPA）对乳腺癌MCF-7细胞中IV型胶原酶（MMP-2、MMP-9）分泌的影响。明胶酶谱分析表明，FGF-2和TPA均刺激MCF-7细胞中MMP-9的分泌，而它们并未改变MMP-2的分泌水平。信号级联研究表明，FGF-2和TPA均诱导Ras激活、c-Raf磷酸化、丝裂原活化蛋白激酶/细胞外信号调节激酶激酶（MEK(1/2)）磷酸化以及细胞外信号调节激酶（ERK(1/2)）磷酸化。用显性负性Ras（Ras-N17）瞬时转染MCF-7细胞以及用MEK(1/2)抑制剂PD98059处理，可显著抑制FGF-2和TPA诱导的MMP-9分泌。发现一种泛蛋白激酶C（PKC）抑制剂GF109203X可完全消除FGF-2和TPA诱导的MMP-9分泌及ERK(1/2)磷酸化。此外，使用如罗特勒素和Gö6976等亚型特异性PKC抑制剂表明，PKC-δ亚型可能是FGF-2和TPA营养信号的一个组成部分。这些结果表明，FGF-2和TPA主要通过PKC依赖的Ras/ERK(1/2)信号通路激活诱导MCF-7细胞中MMP-9的分泌。
