Detection of vascular endothelial growth factor (VEGF) and VEGF receptors Flt-1 and KDR in canine mastocytoma cells
Detection of vascular endothelial growth factor (VEGF) and VEGF receptors Flt-1 and KDR in canine mastocytoma cells
复制标题
DOI：
10.1016/j.vetimm.2006.11.009
复制
发表时间：
2007-02-15
影响因子：
1.8
通讯作者：
Valent, Peter
中科院分区：
文献类型：
--
作者：
Rebuzzi, Laura;Willmann, Michael;Valent, Peter
血管内皮生长因子（VEGF）是血管生成的主要调节因子，也是各种恶性肿瘤中肿瘤细胞的潜在自分泌生长因子。在本研究中，我们研究了VEGF和VEGF受体在犬肥大细胞瘤和犬肥大细胞瘤细胞系C2中的表达。通过组织切片和细胞离心涂片的免疫染色评估，发现原发性肿瘤性肥大细胞（MC）和C2细胞表达VEGF蛋白。在北方印迹和RT-PCR实验中，C2细胞以组成性方式表达VEGF mRNA。LY 294002和雷帕霉素可抑制C2细胞中VEGF mRNA的表达，提示PI 3-kinase/mTOR通路参与其中。C2细胞表达VEGF受体1（Flt-1）和VEGF受体2（KDR）。然而，重组VEGF未能促进C2细胞中的3 H-胸苷摄取，中和抗VEGF抗体（贝伐单抗）未能下调这些细胞中的自发增殖。此外，雷帕霉素在mRNA和蛋白水平上降低C2细胞中VEGF的表达，而不抑制其增殖。总之，犬肥大细胞瘤细胞表达VEGF以及VEGF受体。然而，尽管VEGF及其受体的共表达，VEGF不被犬肥大细胞瘤细胞用作自分泌生长调节剂。(c)2006 Elsevier B. V.保留所有权利。
Vascular endothelial growth factor (VEGF) is a major regulator of angiogenesis and a potential autocrine growth factor for neoplastic cells in various malignancies. In the present study, we have investigated expression of VEGF and VEGF receptors in canine mastocytomas and the canine mastocytoma cell line C2. As assessed by immunostaining of tissue sections and cytospin slides, primary neoplastic mast cells (MC) and C2 cells were found to express the VEGF protein. In Northern blot and RT-PCR experiments, C2 cells expressed VEGF mRNA in a constitutive manner. VEGF mRNA expression in C2 cells was counteracted by LY294002 and rapamycin, suggesting involvement of the PI3-kinase/mTOR pathway. Moreover, C2 cells were found to express VEGF receptor-1 (Flt-1) and VEGF receptor-2 (KDR). However, recombinant VEGF failed to promote 3 H-thymidine uptake in C2 cells, and a neutralizing anti-VEGF antibody (bevacizumab) failed to downregulate spontaneous proliferation in these cells. In addition, rapamycin decreased the expression of VEGF in C2 cells at the mRNA and protein level without suppressing their proliferation. Together, canine mastocytoma cells express VEGF as well as VEGF receptors. However, despite co-expression of VEGF and its receptors, VEGF is not utilized as an autocrine growth regulator by canine mastocytoma cells. (c) 2006 Elsevier B.V. All rights reserved.