Neuropeptide-stimulated cell migration in prostate cancer cells is mediated by RhoA kinase signaling and inhibited by neutral endopeptidase
Neuropeptide-stimulated cell migration in prostate cancer cells is mediated by RhoA kinase signaling and inhibited by neutral endopeptidase
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DOI：
10.1038/sj.onc.1209586
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发表时间：
2006-09
期刊：
影响因子：
8
通讯作者：
Rong Zheng;A. Iwase;R. Shen;O. Goodman;N. Sugimoto;Y. Takuwa;Daniel Lerner;D. Nanus
中科院分区：
文献类型：
--
作者：
Rong Zheng;A. Iwase;R. Shen;O. Goodman;N. Sugimoto;Y. Takuwa;Daniel Lerner;D. Nanus
神经肽蛙皮素和内皮素-1刺激前列腺癌(PC)细胞迁移和侵袭(J Clin Invest，2000；106：1399-1407)。指导这种细胞运动的细胞内信号通路并没有被很好地描绘出来。单体GTP酶RhoA在包括中性粒细胞、单核细胞和成纤维细胞在内的多种细胞中迁移是必需的。我们证明蛙皮素刺激的PC细胞的迁移是通过异源三聚体G蛋白偶联受体(G蛋白)Gα13亚单位激活RhoA和Rho相关的卷曲形成蛋白激酶(ROCK)来实现的。利用小干扰RNA抑制三个已知的G蛋白α亚单位相关的RhoA鸟嘌呤核苷酸交换因子(GEF)的表达，我们还发现其中两个RhoA GEF，PDZ-RhoGf和白血病相关RhoGf(LARG)在PC细胞中以非冗余的方式将蛙皮素受体与RhoA联系起来。接下来，我们展示了激活PDZ-Rhogef和LARG的粘着斑激酶是蛙皮素刺激的RhoA激活所必需的。中性内肽酶(NEP)在正常前列腺上皮细胞中表达，而NEP表达缺失与前列腺癌的进展有关。我们还证明了NEP抑制RhoA的神经肽激活。综上所述，这些结果在PC细胞中建立了从蛙皮素受体到ROCK的连续信号通路，它们暗示NEP是这些细胞中神经肽刺激的RhoA的主要调节因子。这项工作还确定了这一信号通路的成员作为合理的药物操作神经肽刺激的PC细胞迁移的潜在靶点。
The neuropeptides bombesin and endothelin-1 stimulate prostate cancer (PC) cell migration and invasion (J Clin Invest, 2000; 106: 1399–1407). The intracellular signaling pathways that direct this cell movement are not well delineated. The monomeric GTPase RhoA is required for migration in several cell types including neutrophils, monocytes and fibroblasts. We demonstrate that bombesin-stimulated PC cell migration occurs via the heterotrimeric G-protein-coupled receptors (G-protein) Gα13 subunit leading to activation of RhoA, and Rho-associated coiled-coil forming protein kinase (ROCK). Using siRNA to suppress expression of the three known G-protein α-subunit-associated RhoA guanine nucleotide exchange factors (GEFs), we also show that two of these RhoA GEFs, PDZ-RhoGEF and leukemia-associated RhoGEF (LARG), link bombesin receptors to RhoA in a non-redundant manner in PC cells. We next show that focal adhesion kinase, which activates PDZ-RhoGEF and LARG, is required for bombesin-stimulated RhoA activation. Neutral endopeptidase (NEP) is expressed on normal prostate epithelium whereas loss of NEP expression contributes to PC progression. We also demonstrate that NEP inhibits neuropeptide activation of RhoA. Together, these results establish a contiguous signaling pathway from the bombesin receptor to ROCK in PC cells, and they implicate NEP as a major regulator of neuropeptide-stimulated RhoA in these cells. This work also identifies members of this signaling pathway as potential targets for rational pharmacologic manipulation of neuropeptide-stimulated migration of PC cells.