Abstract
The vascular endothelial growth factor (VEGF) is a major mediator of angiogenesis involving tumor growth and metastasis. Polymorphisms in the VEGF gene may regulate VEGF production. In this case–control study, we investigated whether functional polymorphisms (+405 C > G and +936 C > T) in the VEGF gene are associated with the risk of lung cancer. Genomic DNA was isolated from the blood of 100 lung cancer patients and 150 healthy controls, and total RNA was isolated from 48 tumor tissues and adjacent normal lung tissues. Two DNA polymorphisms (+405 C > G and +936 C > T) in the 3′-untranslated regions (3′-UTR) and 5′-untranslated regions (5′-UTR) of vascular endothelial growth factor A (VEGFA) were studied using PCR–RFLP method, and mRNA expression of VEGFA was studied by quantitative real-time PCR. Polymorphisms in the 5′-UTR (+405 C > G) and 3′-UTR (+936 C > T) did not show significant difference between lung cancer cases and control samples (P = 0.11 and P = 0.09, respectively). VEGF +405 CG and GG are significantly more in age group >50 years old, in all grades, and in early pathological stages (P = 0.04, P = 0.03, and P = 0.006, respectively). Also, increased expression of VEGFA mRNA was noted in tumorous compared to non-tumorous tissue (P < 0.0001). Overexpression of the gene was considered at ΔC T > 6.0. Within the group of patients with conventional tumor, those with histology other than squamous cell carcinoma (SCC) had a higher level of VEGFA mRNA expression than SCC patients (P = 0.04). Overexpression of VEGFA mRNA was noted in lung cancer and more so in lung cancer with adenocarcinoma and large cell carcinoma histology and in pathological stages III and IV. VEGFA +405 C > G SNP showed an association with age, pathological grade, and stage.
Similar content being viewed by others
References
Alon, et al. Vascular endothelial growth factor acts as a survival factor for newly formed retinal vessels and has implications for retinopathy of prematurity. Nat Med. 1995;1:1024–8.
Awata, et al. A common polymorphism in the 5′-untranslated region of the VEGF gene is associated with diabetic retinopathy in type 2 diabetes. Diabetes. 2002;51:1635–9.
Balasubramanian, et al. Influence of VEGF-A gene variation and protein levels in breast cancer susceptibility and severity. Int J Cancer. 2007;121:1009–16.
Bell, et al. Oligonucleotide NX1838 inhibits VEGF165-mediated cellular responses in vitro. In Vitro Cell Dev Biol Anim. 1999;35:533–42.
Brem, et al. Prolonged tumor dormancy by prevention of neovascularization in the vitreous. Cancer Res. 1976;36:2807–12.
Cao, et al. Vascular endothelial growth factor C induces angiogenesis in vivo. Proc Natl Acad Sci USA. 1998;95:14389–94.
Connolly, et al. Tumor vascular permeability factor stimulates endothelial cell growth and angiogenesis. J Clin Invest. 1989;84:1470–8.
Ferrara, Davis-Smyth. The biology of vascular endothelial growth factor. Endocr Rev. 1997;18:4–25.
Folkman. Tumor angiogenesis: therapeutic implications. N Engl J Med. 1971;285:1182–6.
Fontanini, et al. Expression of vascular endothelial growth factor mRNA in non-small cell lung carcinomas. Br J Cancer. 1999;79:363–9.
Fontanini, et al. Vascular endothelial growth factors is associated with neovascularization and influences progression of non-small cell lung carcinoma. Clin Cancer Res. 1997;3:861–5.
Formento, et al. Influence of the VEGF-A 936C > T germinal polymorphism on tumoral VEGF expression in head and neck cancer. Pharmacogenomics. 2009;10:1277–83.
Gasparini, et al. Prognostic significance of vascular endothelial growth factor protein in node-negative breast carcinoma. J Natl Cancer Inst. 1997;89:139–47.
Graeven, et al. Melanoma-associated expression of vascular endothelial growth factor and its receptors FLT-1 and KDR. J Cancer Res Clin Oncol. 1999;125:621–9.
Grugel, et al. Both v-Ha-Ras and v-Raf stimulate expression of the vascular endothelial growth factor in NIH 3T3 cells. J Biol Chem. 1995;270:25915–9.
Hanahan, Folkman. Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell. 1996;86:353–64.
Hefler, et al. Vascular endothelial growth factor polymorphisms are associated with prognosis in ovarian cancer. Clin Cancer Res. 2007;13:898–901.
Holmgren, et al. Dormancy of micrometastases: balanced proliferation and apoptosis in the presence of angiogenesis suppression. Nat Med. 1995;1:149–53.
Howell, et al. Influence of vascular endothelial growth factor single nucleotide polymorphisms on tumour development in cutaneous malignant melanoma. Genes Immun. 2002;3:229–32.
Hurwitz, et al. Bevacizumab plus irinotecan, fluorouracil, and leucovorin for metastatic colorectal cancer. N Engl J Med. 2004;350:2335–42.
Jain, et al. The role of vascular endothelial growth factor SNPs as predictive and prognostic markers for major solid tumors. Mol Cancer Ther. 2009;8:2496–508.
Jin, et al. Vascular endothelial growth factor polymorphisms in relation to breast cancer development and prognosis. Clin Cancer Res. 2005;11:3647–53.
Joseph, et al. Androgens regulate vascular endothelial growth factor content in normal and malignant prostatic tissue. Clin Cancer Res. 1997;3:2507–11.
Kabbinavar, et al. Combined analysis of efficacy: the addition of bevacizumab to fluorouracil/leucovorin improves survival for patients with metastatic colorectal cancer. J Clin Oncol. 2005;23:3706–12.
Kataoka, et al. Population-based case–control study of VEGF gene polymorphisms and breast cancer risk among Chinese women. Cancer Epidemiol Biomarkers Prev. 2006;15:1148–52.
Kieser, et al. Mutant p53 potentiates protein kinase C induction of vascular endothelial growth factor expression. Oncogene. 1994;9:963–9.
Koukourakis, et al. VEGF gene sequence variation defines VEGF gene expression status and angiogenic activity in non-small cell lung cancer. Lung Cancer. 2004;46:293–8.
Krippl, et al. A common 936C/T gene polymorphism of vascular endothelial growth factor is associated with decreased breast cancer risk. Int J Cancer. 2003;106:468–71.
Ku, et al. Vascular endothelial growth factor gene −460C/T polymorphism is a biomarker for oral cancer. Oral Oncol. 2005;41:497–502.
Linderholm, et al. Vascular endothelial growth factor is of high prognostic value in node-negative breast carcinoma. J Clin Oncol. 1998;16:3121–8.
Liotta, et al. Quantitative relationships of intravascular tumor cells, tumor vessels, and pulmonary metastases following tumor implantation. Cancer Res. 1974;34:997–1004.
Lu, et al. Association of genetic polymorphisms in the VEGF gene with breast cancer survival. Cancer Res. 2005;65:5015–9.
Michael, et al. VEGF gene sequence variation defines VEGF gene expression status and angiogenic activity in non-small cell lung cancer. Lung Cancer. 2004;46:293–8.
Norrby. Angiogenesis: new aspects relating to its initiation and control. APMIS. 1997;105:417–37.
Ohta, et al. Significance of vascular endothelial growth factor messenger RNA expression in primary lung cancer. Clin Cancer Res. 1996;2:1411–6.
Ohta, et al. Vascular endothelial growth factor and lymph node metastasis in primary lung cancer. Brit J Cancer. 1997;76:1041–5.
Pepper, et al. Angiogenesis: a paradigm for balanced extracellular proteolysis during cell migration and morphogenesis. Enzyme Protein. 1996;49:138–62.
Phillips, et al. Vascular endothelial growth factor (rhVEGF165) stimulates direct angiogenesis in the rabbit cornea. In Vivo. 1994;8:961–5.
Plate, et al. Vascular endothelial growth factor is a potential tumour angiogenesis factor in human gliomas in vivo. Nature. 1992;359:845–8.
Polverini. How the extracellular matrix and macrophages contribute to angiogenesis-dependent diseases. Eur J Cancer. 1996;32A:2430–7.
Rebecca, et al. VEGF polymorphisms and survival in early-stage non-small cell lung cancer. J Clin Oncol. 2008;26(6):856–62.
Renner, et al. A common 936C/T mutation in the gene for vascular endothelial growth factor is associated with vascular endothelial factor plasma levels. J Vasc Res. 2000;37:443–8.
Schneider, et al. Association of vascular endothelial growth factor and vascular endothelial growth factor receptor-2 genetic polymorphisms with outcome in a trial of paclitaxel compared with paclitaxel plus bevacizumab in advanced breast cancer: ECOG 2100. J Clin Oncol. 2008;26:4672–8.
Sfar, et al. Association of VEGF genetic polymorphisms with prostate carcinoma risk and clinical outcome. Cytokine. 2006;35:21–8.
Stevens, et al. Haplotype analysis of the polymorphic human vascular endothelial growth factor gene promoter. Cancer Res. 2003;63:812–6.
Tischer, et al. The human gene for vascular endothelial growth factor. Multiple proteins forms are encoded through alternative exon splicing. J Biol Chem. 1991;266:11947–54.
Tolentino, et al. Vascular endothelial growth factor is sufficient to produce iris neovascularization and neovascular glaucoma in a nonhuman primate. Arch Ophthalmol. 1996;114:964–70.
Torimura, et al. Increased expression of vascular endothelial growth factor is associated with tumor progression in hepatocellular carcinoma. Hum Pathol. 1998;29:986–91.
Tzanakis, et al. Vascular endothelial growth factor polymorphisms in gastric cancer development, prognosis, and survival. J Surg Oncol. 2006;94:624–30.
Vincenti, et al. Assignment of the vascular endothelial growth factor gene to human chromosome 6p21.3. Circulation. 1996;93:1493–5.
Watson, et al. Identification of polymorphisms within the vascular endothelial growth factor (VEGF) gene: correlation with variation in VEGF protein production. Cytokine. 2000;12:1232–5.
Yang, et al. A randomized trial of bevacizumab, an anti-vascular endothelial growth factor antibody, for metastatic renal cancer. N Engl J Med. 2003;349:427–34.
Zhai, et al. Genotypes and haplotypes of VEGF gene are associated with higher ARDS mortality and lower VEGF plasma levels. Thorax. 2007;62:718–22.
Acknowledgement
The authors gratefully acknowledge the financial support provided by the Sher-I-Kashmir Institute of Medical Sciences, Kashmir for this work. Our thanks are also due to the Head and Technical Staff, particularly Miss Shakeela of the operation theater of the Department of Cardiovascular Thoracic Surgery, SKIMS Srinagar who helped us in procuring the tissue samples.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Naik, N.A., Bhat, I.A., Afroze, D. et al. Vascular endothelial growth factor A gene (VEGFA) polymorphisms and expression of VEGFA gene in lung cancer patients of Kashmir Valley (India). Tumor Biol. 33, 833–839 (2012). https://doi.org/10.1007/s13277-011-0306-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13277-011-0306-y