Growth-regulated oncogene is pivotal in thrombin-induced angiogenesis
Growth-regulated oncogene is pivotal in thrombin-induced angiogenesis
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DOI：
10.1158/0008-5472.can-05-2570
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发表时间：
2006-04-15
期刊：
影响因子：
11.2
通讯作者：
Karpatkin, S
中科院分区：
文献类型：
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作者：
Caunt, M;Hu, L;Karpatkin, S
凝血酶诱导的血管生成的机制尚不清楚。使用基因芯片阵列研究凝血酶刺激细胞的促恶性表型，通过 mRNA 和蛋白质分析，我们观察到凝血酶显着上调几种肿瘤细胞系以及内皮细胞中的生长调节癌基因-α (GRO-α)。凝血酶使肿瘤细胞 GRO-α 的分泌增强 25 至 64 倍。 GRO-alpha 是一种 CXC 趋化因子，在连接其 CXCR2 受体后具有肿瘤相关的血管生成和致癌激活作用。在鸡绒毛尿囊膜测定中，GRO-α 使血管生成增强 2.2 倍，为 GRO-α 作为血管生成生长因子提供了直接证据。抗 GRO-α 抗体完全抑制凝血酶诱导的血管生成上调 2.7 倍，以及基质胶中内皮细胞索形成和体外生长的凝血酶诱导上调 1.5 倍。凝血酶及其PARA受体激活肽[凝血酶受体激活肽(TRAP)]以及GRO-α均显着增加人脐静脉内皮细胞中的血管调节蛋白和生长因子：基质金属蛋白酶(MMP)-1、MMP-2、血管内皮生长因子(VEGF)、血管生成素-2(Ang-2)、CD31以及受体KDR和CXCR2。所有凝血酶/TRAP基因上调均被抗GRO-α抗体完全抑制，并且不受不相关抗体的影响。在小干扰 RNA (shRNA) GRO-α KD 4TI 乳腺肿瘤和 B16F10 黑色素瘤细胞中发现了类似的基因上调抑制以及凝血酶诱导的趋化性。使用 shRNA GRO-a 对野生型小鼠进行体内肿瘤生长研究。 KD 细胞显示肿瘤生长、转移和血管生成受损 2 至 4 倍，而不受内源性凝血酶的影响。因此，凝血酶诱导的血管生成需要GRO-a的上调。肿瘤细胞和内皮细胞中 GRO-α 的凝血酶上调有助于肿瘤血管生成。
The mechanism of thrombin-induced angiogenesis is poorly understood. Using a gene chip array to investigate the promalignant phenotype of thrombin-stimulated cells, we observed that thrombin markedly up-regulates growth-regulated oncogene-alpha (GRO-alpha) in several tumor cell lines as well as endothelial cells by mRNA and protein analysis. Thrombin enhanced the secretion of GRO-alpha from tumor cells 25- to 64fold. GRO-alpha is a CXC chemokine with tumor-associated angiogenic as well as oncogenic activation following ligation of its CXCR2 receptor. GRO-alpha enhanced angiogenesis in the chick chorioallantoic membrane assay 2.2-fold, providing direct evidence for GRO-alpha as an angiogenic growth factor. Anti-GRO-alpha antibody completely inhibited the 2.7-fold thrombin-induced up-regulation of angiogenesis, as well as the 1.5-fold thrombin-induced up-regulation of both endothelial cell cord formation in Matrigel and growth in vitro. Thrombin as well as its PARA receptor activation peptide [thrombin receptor activation peptide (TRAP)] as well as GRO-alpha all markedly increased vascular regulatory proteins and growth factors: matrix metalloproteinase (MMP)-1, MMP-2, vascular endothelial growth factor (VEGF), angiopoictin-2 (Ang-2), CD31, and receptors KDR and CXCR2 in human umbilical vein endothelial cells. All of the thrombin/TRAP gene upregulations were completely inhibited by anti-GRO-alpha antibody and unaffected by irrelevant antibody. Similar inhibition of gene up-regulation as well as thrombin-induced chemotaxis was noted with small interfering RNA (shRNA) GRO-alpha KD 4TI breast tumor and B16F10 melanoma cells. In vivo tumor growth studies in wild-type mice with shRNA GRO-a. KD cells revealed 2- to 4-fold impaired tumor growth, metastasis, and angiogenesis, which was not affected by endogenous thrombin. Thus, thrombin-induced angiogenesis requires the up-regulation of GRO-a. Thrombin up-regulation of GRO-alpha in tumor cells as well as endothelial cells contributes to tumor angiogenesis.