VEGF polymorphisms are not associated with an increased risk of developing renal cell carcinoma in Spanish population Pablo Sáenz-López a,c,1 , Fernando Vazquez b,1 , Jose Manuel Cozar b , Rafael Carretero a,c , Federico Garrido a,c , Francisco Ruiz-Cabello a,c,⇑ a Departamento de Bioquímica III e Inmunología, Universidad de Granada, Spain b Servicio de Urología, Hospital Universitario Virgen de las Nieves, Granada, Spain c Servicio de Análisis Clínicos e Inmunología, Hospital Universitario Virgen de las Nieves, Granada, Spain article info Article history: Received 25 June 2012 Accepted 3 October 2012 Available online 13 October 2012 abstract Purpose: Vascular endothelial growth factor (VEGF) plays a central role in promoting angiogenesis and is over-expressed in renal cell cancer (RCC). Published data on the association between polymorphisms of vascular endothelial growth factor (e.g., À2578C/A [rs699947], À460T/C [rs833061], +405C/G [rs2010 963], and +936C/T [rs3025039]) and the risk of renal cell carcinoma are ambiguous and controversial. The aim of this investigation was to investigate this relationship in a series of Caucasian Spanish patients. Materials and methods: A case-control study was performed with 216 cases and 280 controls, genotyping subjects for VEGF polymorphisms using the predesigned TaqMan single nucleotide polymorphism (SNP) genotyping assay (Applied Biosystems, Foster City, CA, USA). The combined effect of the four gene poly- morphisms on overall survival was studied by haplotype analysis. Results: The overall results suggest that polymorphisms or haplotypes in the VEGF gene do not modify the risk of RCC. We were unable to replicate the association of the À460T/C (rs833061) polymorphism with renal cancer risk. Data were also gathered on clinical-pathological results, tumor size, clinical stage, histological grade, and survival. Conclusions: According to our analysis of their contribution to prognostic factors, VEGF polymorphisms do not appear to exert a significant influence on RCC progression or prognosis. This finding might be explained by the tumor biology and pathogenesis of clear cell RCC. Additional studies with larger sample sizes are needed in different ethnic groups to further assess this association. Ó 2012 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved. 1. Introduction Angiogenesis is a relatively early event in carcinogenesis [1]. Newly formed blood vessels are formed from endothelial precur- sors and play a key role in cancer growth, dispose of the metabolic waste products of tumor cells, supply the tumor with nutrients and oxygen, generate paracrine stimuli, and provide potential routes for tumor progression and metastasis [2]. Angiogenesis is regulated by various growth factors, and a critical role is played by the vascular endothelial growth factor (VEGF or VEGF-A), a soluble 34–46 kDa heparin-binding glycoprotein dimmer. It specifically binds to different VEGF receptors (VEGFRs) that belong to the tyrosine-kinase family of receptors on endothelial cells and initiate intracellular signal transduction pathways mediating angiogenesis and vascular permeability [3]. Several experiments found in- creased VEGF expression to be associated with tumor growth and metastasis, whereas the inhibition of VEGF signaling suppressed tumor-induced angiogenesis and tumor growth [4]. The gene encoding VEGF is located on chromosome 6 and comprises a 14-kb coding region with 8 exons and 7 introns [5]. At least five different isoforms of VEGF are generated by alternative splicing of the VEGF gene [4]. VEGF production has been identified in numerous cells, including macrophages, keratinocytes, megakary- ocytes, T cells, smooth muscle cells, basophiles, fibroblasts, mast cells, and neutrophils, and VEGF is expressed in brain, kidney, liver, and spleen, among other tissues [6]. VEGF gene expression is regulated by various effectors, including lipopolysaccharides (LPS), cytokines (e.g., IL-6), hormones (e.g., luteotrophic hormone), and growth factors (PDGF, EGF) as well as by hypoxia [4]. The VEGF is a highly polymorphic gene, and a number of single nucleotide polymorphisms (SNPs) have been reported [7]. Polymorphisms in its promoter region (loci À2578C/A [rs699947] and À460T/C [rs833061]), 5 0 -untranslated region (+405C/G [rs201 0963]), and 3 0 -untranslated region (+936C/T [rs3025039]) have been 0198-8859/$36.00 - see front matter Ó 2012 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.humimm.2012.10.014 ⇑ Corresponding author at: Hospital Universitario Virgen de las Nieves, Avenida de las Fuerzas Armadas s/n, 18014 Granada, Spain. Fax: +34 958020120. E-mail address: fruizc@ugr.es (F. Ruiz-Cabello). 1 The authors Pablo Sáenz-López and Fernando Vazquez have equally contributed to this work and the two should be considered as first authors. Human Immunology 74 (2013) 98–103 Contents lists available at SciVerse ScienceDirect www.ashi-hla.org journal homepage: www.elsevier.com/locate/humimm