Hepatocyte growth factor-mediated cell invasion in pancreatic cancer cells is dependent on neuropilin-1
Hepatocyte growth factor-mediated cell invasion in pancreatic cancer cells is dependent on neuropilin-1
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DOI：
10.1158/0008-5472.can-07-3256
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发表时间：
2007-11-01
期刊：
影响因子：
11.2
通讯作者：
Korc, Murray
中科院分区：
文献类型：
--
作者：
Matsushita, Arikira;Gotze, Tobias;Korc, Murray
神经匹林-1 （Neuropilin-1, Np-1）是信号蛋白3A和血管内皮生长因子的受体，在胰腺导管腺癌（PDAC）中高水平表达。为了评估Np-1在PDAC中的潜在作用，将表达相对较低水平Np-1的COLO-357胰腺癌细胞稳定转染Np-1 cDNA。Np-1过表达与肝细胞生长因子（HGF）对细胞侵袭性的增强有关，而这种作用被小干扰rna介导的c-Met下调所消除。相反，在Np-1表达水平相对较高的PANC-1胰腺癌细胞中，抑制内源性Np-1可完全消除hgf介导的细胞侵袭。为了确定哪些途径参与了np -1介导的c- met依赖性细胞侵袭性的促进，接下来在假转染和np -1过表达的COLO-357细胞中检测了HGF对信号传导的影响。与假转染的细胞相比，在np -1过表达的COLO-357细胞中，HGF对c-Met酪氨酸磷酸化和p38丝裂原活化蛋白激酶（MAPK）活化的作用增加。p38 MAPK抑制剂SB203580可以抑制np -1过表达细胞中hgf诱导的侵袭，而MAP/细胞外信号调节激酶抑制剂U0126则没有作用。Src抑制剂PP2和磷脂酰肌醇3-激酶抑制剂LY294002也能抑制hgf诱导的细胞侵袭。免疫沉淀研究表明，Np-1与c-Met相关，但与表皮生长因子受体家族成员无关。共聚焦显微镜显示，这种关联发生在质膜上，HGF促进了Np-1-c-Met复合物的内化，导致其在核周定位。这些发现表明，Np-1是有效激活促进细胞侵袭的c- met依赖途径所必需的。
Neuropilin-1 (Np-1), a receptor for semaphorin 3A and vascular endothelial growth factor, is expressed at high levels in pancreatic ductal adenocarcinoma (PDAC). To assess the potential role of Np-1 in PDAC, COLO-357 pancreatic cancer cells, which express relatively low levels of Np-1, were stably transfected with the Np-1 cDNA. Np-1 overexpression was associated with enhanced cell invasiveness in response to hepatocyte growth factor (HGF), and this effect was abolished by small interfering RNA-mediated down-regulation of c-Met. Conversely, in PANC-1 pancreatic cancer cells, which express relatively high levels of Np-1, suppression of endogenous Np-1 completely abolished HGF-mediated cell invasion. To determine which pathways are involved in Np-1-mediated facilitation of c-Met-dependent cell invasiveness, the effects of HGF on signaling were examined next in sham-transfected and Np-1-overexpressing COLO-357 cells. HGF actions on c-Met tyrosine phosphorylation and p38 mitogen-activated protein kinase (MAPK) activation were increased in Np-1-overexpressing COLO-357 cells by comparison with HGF effects in sham-transfected cells. SB203580, an inhibitor of p38 MAPK, suppressed HGF-induced invasion in Np-1-overexpressing cells, whereas U0126, a MAP/extracellular signal-regulated kinase kinase inhibitor, was without effect. PP2, a Src inhibitor, and LY294002, a phosphatidylinositol 3-kinase inhibitor, also suppressed HGF-induced invasion in these cells. Immunoprecipitation studies revealed that Np-1 associated with c-Met, but not with epidermal growth factor receptor, family members. Confocal microscopy indicated that this association occurred on the plasma membrane and that HGF promoted the internalization of Np-1-c-Met complex, leading to its perinuclear localization. These findings indicate that Np-1 is required for efficient activation of c-Met-dependent pathways that promote cell invasiveness.