Identification of a natural soluble neuropilin-1 that binds vascular endothelial growth factor:: In vivo expression and antitumor activity
Identification of a natural soluble neuropilin-1 that binds vascular endothelial growth factor:: In vivo expression and antitumor activity
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DOI：
10.1073/pnas.040337597
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发表时间：
2000-03-14
影响因子：
11.1
通讯作者：
Klagsbrun, M
中科院分区：
文献类型：
--
作者：
Gagnon, ML;Bielenberg, DR;Klagsbrun, M
神经纤毛蛋白-1（NRP 1）是一种130-kDa的跨膜受体，用于脑信号蛋白（神经元导向介质）和血管内皮生长因子165（VEGF（165））（血管生成因子）。克隆了一个2.2 kb截短的NRP 1 cDNA，其编码一个644-aa的可溶性NRP 1（sNRP 1）同种型，仅含有NRP 1的a/CU B和B/凝血因子同源性胞外结构域。sNRP 1由细胞分泌为结合VECF 165的90-kDa蛋白。而不是VEGF（121）。它抑制I-125-VEGF（165）与内皮细胞和肿瘤细胞的结合以及VEGF（165）诱导的内皮细胞中KDR的酪氨酸磷酸化。sNRP 1cDNA的3'端含有一个独特的28 bp内含子衍生序列，该序列在全长NRP 1cDNA中是不存在的。使用对应于该独特序列的探针，sNRP 1 mRNA可以通过原位杂交与全长NRP 1 mRNA进行差异检测，例如，在肝、肾、皮肤和乳腺细胞中。血管原位分析显示NRP 1而非sNRP 1表达。sNRP 1在体内有功能。与对照肿瘤不同，表达重组sNRP 1的大鼠前列腺癌细胞肿瘤的特征在于广泛出血。受损血管和凋亡肿瘤细胞。这些结果表明存在天然存在的可溶性NRP 1，其表达不同于完整的NRP 1，并且似乎是VEGF 165拮抗剂。
Neuropilin-1 (NRP1) is a 130-kDa transmembrane receptor for semaphorins, mediators of neuronal guidance, and for vascular endothelial growth factor 165 (VEGF(165)), an angiogenesis factor. A 2.2-kb truncated NRP1 cDNA was cloned that encodes a 644-aa soluble NRP1 (sNRP1) isoform containing just the a/CUB and b/coagulation factor homology extracellular domains of NRP1. sNRP1 is secreted by cells as a 90-kDa protein that binds VECF165. but not VEGF(121). It inhibits I-125-VEGF(165) binding to endothelial and tumor cells and VEGF(165)-induced tyrosine phosphorylation of KDR in endothelial cells. The 3' end of sNRP1 cDNA contains a unique, 28-bp intron-derived sequence that is absent in full-length NRP1 cDNA, Using a probe corresponding to this unique sequence, sNRP1 mRNA could be detected by in situ hybridization differentially from full-length NRP1 mRNA, for example, in cells of liver, kidney, skin, and breast. Analysis of blood vessels in situ showed that NRP1, but not sNRP1, was expressed. sNRP1 was functional in vivo. Unlike control tumors, tumors of rat prostate carcinoma cells expressing recombinant sNRP1 were characterized by extensive hemorrhage. damaged vessels, and apoptotic tumor cells. These results demonstrate the existence of a naturally occurring, soluble NRP1 that is expressed differently from intact NRP1 and that appears to be a VEGF165 antagonist.