Suppression of vascular endothelial growth factor receptor 3 (VEGFR3) and vascular endothelial growth factor C (VEGFC) inhibits hypoxia-induced lymph node metastases in cervix cancer
Suppression of vascular endothelial growth factor receptor 3 (VEGFR3) and vascular endothelial growth factor C (VEGFC) inhibits hypoxia-induced lymph node metastases in cervix cancer
复制标题
DOI：
10.1016/j.ygyno.2011.07.006
复制
发表时间：
2011-11-01
影响因子：
4.7
通讯作者：
Hill, Richard P.
中科院分区：
文献类型：
--
作者：
Chaudary, Naz;Milosevic, Michael;Hill, Richard P.
目标.我们已经研究了暴露于缺氧后VEGFC/VEGFR 3信号传导在原位人ME 180和SiHa宫颈异种移植模型的淋巴结转移和生长中的作用。我们以前的研究表明，这些肿瘤的生长条件下的循环缺氧增加淋巴结转移。荷瘤小鼠接受7%O-2/空气10 min循环缺氧，4 h/天/2周。通过shRNA进行vegfc的敲低，并且通过针对小鼠和人蛋白质的阻断抗体进行VEGFR 3的抑制。Western blotting检测VEGF 3蛋白表达。免疫组织化学染色检测CA 9、CD 31、LYVE 1和Ki 67的表达。通过真实的时间PCR测定基因表达。在常氧和循环缺氧条件下，敲低vegfc或用阻断抗体抑制VEGFR 3可减少转移。在vegfc敲低和VEGFR 3抑制后，观察到血管生成和血管形成的减少以及肿瘤细胞增殖的减少。低氧后VEGFR 3在mRNA和蛋白水平上表达上调。总的来说，我们的结果表明，抗VEGFR 3抗体抑制vegfc诱导的肿瘤淋巴管生成和转移，并且vegfc在肿瘤细胞中的敲低引起对淋巴结转移的类似抑制作用。这些结果表明，vegfc/NEGFR 3对肿瘤细胞形成淋巴结转移的进展的影响主要发生在低氧肿瘤微环境下。(C)2011 Elsevier Inc. All rights reserved.
Objectives. We have examined the role of VEGFC/VEGFR3 signaling in lymph node metastasis and growth of orthotopic human ME180 and SiHa cervical xenograft models following exposure to hypoxia. Our previous studies showed that growth of these tumors under conditions of cyclic hypoxia increased nodal metastasis.Methods. Mice bearing orthotopic tumors were subjected to cyclic hypoxia at 7% O-2/air 10 min cycles 4 h/day/2 weeks. Knockdown of vegfc was carried out by shRNA and inhibition of VEGFR3 was conducted by blocking antibodies for the mouse and human proteins. VEGFR3 protein expression was detected by Western blotting. Immunohistochemical staining was used to assess CA9, CD31, LYVE1 and Ki67 labeling. Gene expression was determined by real time PCR.Results. Knock down of vegfc or inhibition of VEGFR3 with blocking antibody reduced metastases under normoxic and cyclic hypoxia conditions. A reduction in lymphatics and blood vessel formation and a decrease in tumor cell proliferation was observed following vegfc knockdown and VEGFR3 inhibition. VEGFR3 expression was upregulated at the mRNA and protein levels following hypoxia.Conclusions. Collectively, our results indicate that anti-VEGFR3 antibody inhibits vegfc-induced tumor lymphangiogenesis and metastasis and that vegfc knockdown in the tumor cells causes a similar inhibitory effect on lymph node metastasis. These results suggest that the effects of vegfc/NEGFR3 on the progression of tumor cells to form lymph node metastases occur primarily under an hypoxic tumor microenvironment. (C) 2011 Elsevier Inc. All rights reserved.