ORIGINAL PAPER Vascular endothelial growth factor gene 936 C/T polymorphism in breast cancer patients Aydan Erog ˘lu Æ Ays ¸enur O ¨ ztu ¨rk Æ Ragıp C ¸ am Æ Nejat Akar Received: 30 March 2007 / Accepted: 22 June 2007 / Published online: 1 August 2007 Ó Humana Press Inc. 2007 Angiogenesis is an important step in the development, growth, and metastasis of malignant tumor. Vascular endothelial growth factor (VEGF) is one of the most potent endothelial cell mitogens and plays an important role in angiogenesis. High level of VEGF and increased micro- vessel density in tumors are associated with an advanced stage disease and worse prognosis for several types of malignant tumors, including breast cancer [1, 2]. The VEGF gene is located on chromosome 6p21.3 and consist 8 exons [3]. Several polymorphisms have been described in the VEGF gene and they have been associated with increased risk of several tumors [4–10]. One of the most common single nucleotide polymor- phisms is C+936T in the 3¢-untranslated region and this polymorphism is related to VEGF protein production [11]. Studies on the association of 936 C/T polymor- phism of the VEGF gene with breast cancer risk have conflicting findings [4–7]. We have recently reported that 936 C/T polymorphism of VEGF gene is related to the development of various types cancers [12]. In this study, we have investigated the association of this polymor- phism with breast cancer risk and clinico-pathological characteristics of the disease. We enrolled two groups. The first group was composed 60 patients with invasive breast cancer (mean age 51.2 ± 12 years; range 23–77 years). Patient and tumor characteristics were recorded. The control group consisted of 60 unrelated age-matched healthy volunteer women without history of malignancy. Genomic DNA isolation was performed from peripheral venous blood by standard phenol–chlorophorm extraction, and polymerase chain reaction of VEGF C936T polymor- phism was performed according to previously described method [11] using forward primer 5¢-AAG GAA GAG ACT CTG CGC-3¢ and reverse primer 5¢-TAT GTG GGT GGG TGT GTC TAC AGG-3¢. Amplification was per- formed for 34-cycles with an annealing temperature of 60°C (Biometra, Germany). Amplified DNA was subjected to 2.5% agarose gel electrophoresis. The statistical analyses were performed using SPSS software (SPSS Inc., Chicago, IL). The chi-squared test or Fischer’s exact test was used to detect difference in the genotype frequencies between the groups. Value of P < 0.05 was considered to be statistically significant. No 936 TT genotype of VEGF gene was detected in both patient and control groups. Carriers of 936 CT genotype were more frequent among patient group (16 of 60 patients, 26.7%) than among control group (3 of 60 cases, 5%) and this difference reached statistical signifi- cance (P = 0.001). The associations of VEGF 936 geno- type with clinicopathological characteristics were listed in Table 1. No correlation was found between VEGF 936 C/T polymorphism and the characteristics. Recent studies have shown that 936 C/T polymorphism of the VEGF gene is not significantly associated with invasive breast cancer [6, 7]. In contrast, Krippl et al. [4] reported that carriers of CT or TT genotype were correlated A. Erog ˘lu (&) Department of General Surgery and Surgical Oncology, Numune State Hospital, Konya, Turkey e-mails: aydaneroglu@hotmail.com; aydaneroglu@gmail.com A. O ¨ ztu ¨rk Á N. Akar Department of Pediatric Molecular Genetic, Ankara University Medical School, Ankara, Turkey R. C ¸ am Department of General Surgery, Ankara University Medical School, Ankara, Turkey Med Oncol (2008) 25:54–55 DOI 10.1007/s12032-007-0046-4