Role of vascular endothelial growth factor in non-small cell lung cancer pathogenesis

The angiogenesis is an important process in the pathogenesis of malignancies. It is regulated by various growth factors, with the vascular endothelial growth factor (VEGF) playing the central role. The aim of the present study was to evaluate possible associations of functional VEGF −2578C>A, −63...

Full description

Saved in:
Bibliographic Details
Published in:Experimental Oncology
Date:2015
Main Authors: Krupnova, E.V., Shapetska, M.N., Mikhalenko, E.P., Chebotaryova, N.V., Shchayuk, A.N., Pissarchik, S.N., Prokhorov, A.V.
Format: Article
Language:English
Published: Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України 2015
Subjects:
Online Access:https://nasplib.isofts.kiev.ua/handle/123456789/145534
Tags: Add Tag
No Tags, Be the first to tag this record!
Journal Title:Digital Library of Periodicals of National Academy of Sciences of Ukraine
Cite this:Role of vascular endothelial growth factor in non-small cell lung cancer pathogenesis / E.V. Krupnova, M.N. Shapetska, E.P. Mikhalenko, N.V. Chebotaryova, A.N. Shchayuk, S.N. Pissarchik, A.V. Prokhorov // Experimental Oncology. — 2015. — Т. 37, № 3. — С. 213-217. — Бібліогр.: 38 назв. — англ.

Institution

Digital Library of Periodicals of National Academy of Sciences of Ukraine
id nasplib_isofts_kiev_ua-123456789-145534
record_format dspace
spelling Krupnova, E.V.
Shapetska, M.N.
Mikhalenko, E.P.
Chebotaryova, N.V.
Shchayuk, A.N.
Pissarchik, S.N.
Prokhorov, A.V.
2019-01-22T19:49:12Z
2019-01-22T19:49:12Z
2015
Role of vascular endothelial growth factor in non-small cell lung cancer pathogenesis / E.V. Krupnova, M.N. Shapetska, E.P. Mikhalenko, N.V. Chebotaryova, A.N. Shchayuk, S.N. Pissarchik, A.V. Prokhorov // Experimental Oncology. — 2015. — Т. 37, № 3. — С. 213-217. — Бібліогр.: 38 назв. — англ.
1812-9269
https://nasplib.isofts.kiev.ua/handle/123456789/145534
The angiogenesis is an important process in the pathogenesis of malignancies. It is regulated by various growth factors, with the vascular endothelial growth factor (VEGF) playing the central role. The aim of the present study was to evaluate possible associations of functional VEGF −2578C>A, −634G>C, and +936C>T polymorphisms with the risk for occurrence and progression of non-small cell lung cancer (NSCLC) in patients living in Republic of Belarus. Materials and Methods: A total of 202 patients (147 males and 55 females) diagnosed as having the NSCLC. The control group consisted of 336 individuals (245 males and 91 females) without an oncopathology. The total DNA was isolated from peripheral blood. We investigated the single nucleotide polymorphisms of VEGF (rs 2010963), (rs 699947), (rs 3025039). The genotyping was performed by PCR-RFLP analysis. Results: Our results revealed a marginally significant association of the –2578CC genotype (p=0.002) with a greater degree of tumor spread (Т2–Т4). Heterozygous genotypes –2578СА and +936СT carriers were included into the follow-up group significantly more often (р=0.021 and р=0.012, respectively). Our study demonstrate that VEGF –2578A/C and +936C/T polymorphisms are among the factors determining the individual peculiarities of NSCLC course in this population and can be used for clarifying the prognosis of the disease. Key Words: vascular endothelial growth factor, polymorphism, non-small cell lung cancer.
en
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
Experimental Oncology
Original contributions
Role of vascular endothelial growth factor in non-small cell lung cancer pathogenesis
Article
published earlier
institution Digital Library of Periodicals of National Academy of Sciences of Ukraine
collection DSpace DC
title Role of vascular endothelial growth factor in non-small cell lung cancer pathogenesis
spellingShingle Role of vascular endothelial growth factor in non-small cell lung cancer pathogenesis
Krupnova, E.V.
Shapetska, M.N.
Mikhalenko, E.P.
Chebotaryova, N.V.
Shchayuk, A.N.
Pissarchik, S.N.
Prokhorov, A.V.
Original contributions
title_short Role of vascular endothelial growth factor in non-small cell lung cancer pathogenesis
title_full Role of vascular endothelial growth factor in non-small cell lung cancer pathogenesis
title_fullStr Role of vascular endothelial growth factor in non-small cell lung cancer pathogenesis
title_full_unstemmed Role of vascular endothelial growth factor in non-small cell lung cancer pathogenesis
title_sort role of vascular endothelial growth factor in non-small cell lung cancer pathogenesis
author Krupnova, E.V.
Shapetska, M.N.
Mikhalenko, E.P.
Chebotaryova, N.V.
Shchayuk, A.N.
Pissarchik, S.N.
Prokhorov, A.V.
author_facet Krupnova, E.V.
Shapetska, M.N.
Mikhalenko, E.P.
Chebotaryova, N.V.
Shchayuk, A.N.
Pissarchik, S.N.
Prokhorov, A.V.
topic Original contributions
topic_facet Original contributions
publishDate 2015
language English
container_title Experimental Oncology
publisher Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
format Article
description The angiogenesis is an important process in the pathogenesis of malignancies. It is regulated by various growth factors, with the vascular endothelial growth factor (VEGF) playing the central role. The aim of the present study was to evaluate possible associations of functional VEGF −2578C>A, −634G>C, and +936C>T polymorphisms with the risk for occurrence and progression of non-small cell lung cancer (NSCLC) in patients living in Republic of Belarus. Materials and Methods: A total of 202 patients (147 males and 55 females) diagnosed as having the NSCLC. The control group consisted of 336 individuals (245 males and 91 females) without an oncopathology. The total DNA was isolated from peripheral blood. We investigated the single nucleotide polymorphisms of VEGF (rs 2010963), (rs 699947), (rs 3025039). The genotyping was performed by PCR-RFLP analysis. Results: Our results revealed a marginally significant association of the –2578CC genotype (p=0.002) with a greater degree of tumor spread (Т2–Т4). Heterozygous genotypes –2578СА and +936СT carriers were included into the follow-up group significantly more often (р=0.021 and р=0.012, respectively). Our study demonstrate that VEGF –2578A/C and +936C/T polymorphisms are among the factors determining the individual peculiarities of NSCLC course in this population and can be used for clarifying the prognosis of the disease. Key Words: vascular endothelial growth factor, polymorphism, non-small cell lung cancer.
issn 1812-9269
url https://nasplib.isofts.kiev.ua/handle/123456789/145534
citation_txt Role of vascular endothelial growth factor in non-small cell lung cancer pathogenesis / E.V. Krupnova, M.N. Shapetska, E.P. Mikhalenko, N.V. Chebotaryova, A.N. Shchayuk, S.N. Pissarchik, A.V. Prokhorov // Experimental Oncology. — 2015. — Т. 37, № 3. — С. 213-217. — Бібліогр.: 38 назв. — англ.
work_keys_str_mv AT krupnovaev roleofvascularendothelialgrowthfactorinnonsmallcelllungcancerpathogenesis
AT shapetskamn roleofvascularendothelialgrowthfactorinnonsmallcelllungcancerpathogenesis
AT mikhalenkoep roleofvascularendothelialgrowthfactorinnonsmallcelllungcancerpathogenesis
AT chebotaryovanv roleofvascularendothelialgrowthfactorinnonsmallcelllungcancerpathogenesis
AT shchayukan roleofvascularendothelialgrowthfactorinnonsmallcelllungcancerpathogenesis
AT pissarchiksn roleofvascularendothelialgrowthfactorinnonsmallcelllungcancerpathogenesis
AT prokhorovav roleofvascularendothelialgrowthfactorinnonsmallcelllungcancerpathogenesis
first_indexed 2025-11-24T20:37:53Z
last_indexed 2025-11-24T20:37:53Z
_version_ 1850492878181105664
fulltext Experimental Oncology 37, 213–217, 2015 (September) 213 ROLE OF VASCULAR ENDOTHELIAL GROWTH FACTOR IN NON-SMALL CELL LUNG CANCER PATHOGENESIS E.V. Krupnova1,*, M.N. Shapetska2, E.P. Mikhalenko1, N.V. Chebotaryova1, A.N. Shchayuk1, S.N. Pissarchik3, A.V. Prokhorov2 1Institute of Genetics and Cytology, NAS of Belarus, Minsk 220072, Republic of Belarus 2Belarusian State Medical University, Minsk 220116, Republic of Belarus 3City Clinical Pathologoanatomic Bureau, Minsk 220116, Republic of Belarus The angiogenesis is an important process in the pathogenesis of malignancies. It is regulated by various growth factors, with the vas- cular endothelial growth factor (VEGF) playing the central role. The aim of the present study was to evaluate possible associations of functional VEGF −2578C>A, −634G>C, and +936C>T polymorphisms with the risk for occurrence and progression of non-small cell lung cancer (NSCLC) in patients living in Republic of Belarus. Materials and Methods: A total of 202 patients (147 males and 55 females) diagnosed as having the NSCLC. The control group consisted of 336 individuals (245 males and 91 females) without an oncopathology. The total DNA was isolated from peripheral blood. We investigated the single nucleotide polymorphisms of VEGF (rs 2010963), (rs 699947), (rs 3025039). The genotyping was performed by PCR-RFLP analysis. Results: Our results revealed a marginally significant association of the –2578CC genotype (p=0.002) with a greater degree of tumor spread (Т2–Т4). Heterozy- gous genotypes –2578СА and +936СT carriers were included into the follow-up group significantly more often (р=0.021 and р=0.012, respectively). Our study demonstrate that VEGF –2578A/C and +936C/T polymorphisms are among the factors determining the in- dividual peculiarities of NSCLC course in this population and can be used for clarifying the prognosis of the disease. Key Words: vascular endothelial growth factor, polymorphism, non-small cell lung cancer. The angiogenesis is an important process in the pa- t hogenesis of malignancies. It is regulated by various growth factors, with the vascular endothelial growth factor (VEGF-A or VEGF) playing the central role [1]. The VEGF gene expression is closely related to the de- gree of vascularization and the prognosis for the oc- currence of numerous solid tumors and is considered to be a predictor of resistance to the chemo- and ra- diotherapy [2]. An elevated VEGF expression is associ- ated with tumor growth and metastatic process, while the inhibited VEGF expression results in suppressed tumor growth [3]. The VEGF gene also triggers the activation of the pro- tease cascade involved in the degradation of extracellular matrix, suppressing apoptosis, stimulates the endothelial cells survival, increases vascular permeability, inhibits the dendritic cells differentiation, regulates the hexose transport in endothelial cells as well as activates tissue factors and monocytes migration [1]. Clinical studies have shown that the high level of VEGF expression and, respectively, the increased number of microvessels in the tumor correlate with the disease stage and the un- favorable prognosis for many tumor types, including the non-small cell lung cancer (NSCLC) [4–8]. The VEGF gene is located on the short arm of chromosome 6 (6p21.3) and consists of eight exons and seven introns [7]. VEGF is a diametric glycoprotein, acting via tyrosine kinase receptors VEGFR1 and VEGFR2 located predominantly on en- dothelial cells [9]. Polymorphic sites of VEGF gene determining the level of VEGF production are found in the promoter — 5´-un- translated region (5´-UTR) as well as in 5´- and 3´-un- translated regions of the gene (3´-UTR). The VEGF gene polymorphism is associated with differentiated VEGF expression and protein produc- tion. VEGF-2578C/C, −634 C/-genotypes are related to high level of VEGF expression [5, 9–12], while +936T allele correlates with low VEGF expression and its low levels in blood plasma [13, 14]. Recent studies show that individual polymorphic variants of VEGF influence the development of cer- tain cancers. In particular, VEGF C-634G, C-2578A, C+936T polymorphisms are associated with an in- creased lung cancer risk in Asian population [15]. Based on the above-mentioned, it can be assumed that these single nucleotide substitutions of the VEGF gene related to angiogenesis can affect the risk of tumour occur- rence and progression as well as the patients’ survival. The aim of the present study was to evaluate pos- sible associations of functional VEGF −2578C>A, −634G>C, and +936C>T polymorphisms with the risk for occurrence and progression of NSCLC in patients living in the territory of the Republic of Belarus. MATERIALS AND METHODS 202 patients (147 males and 55 females) diag- nosed as having the NSCLC and treated at the Minsk City Oncology Dispensary during the period from 2003 to 2012 were included in the study. The control group consisted of 336 individuals (245 males and 91 females) without an oncopathology who were age-, gender- and comorbidity matched with NSCLC pa- Submitted: January 20, 2015. *Correspondence: E-mail: ekrupnova@inbox.ru Fax: (+375 17) 284-19-17 Abbreviations used: 3´-UTR — 3´-untranslated region; 5´-UTR — 5´-untranslated region; NSCLC — non-small cell lung cancer; VEGF — vascular endothelial growth factor. Exp Oncol 2015 37, 3, 213–217 214 Experimental Oncology 37, 213–217, 2015 (September) tients. Clinical characteristics of patients with NSCLC and controls are presented in Table 1. Table 1. Characteristics of study population Characteristics Patients with NSCLC (n = 202) Controls (n = 336) n (%) n (%) Gender: female 55 (27.2) 91 (27.1) male 147 (72.8) 245 (72.9) Smoking status: smokes 67 (33.2) 201 (59.8) does not smoke 119 (58.9) 135 (40.2) no information 16 (7.9) − Stage: I 108 (53.5) − II 27 (13.4) − III 55 (27.2) − IV 12 (5.9) − Histology: squamous-cell carcinoma 106 (52.5) − adenocarcinoma 96 (47.5) − Surgery: lobectomy/bilobectomy 121 (65.1) − pneumonectomy 42 (22.6) − lung resection 13 (7.0) − biopsy 10 (5.4) − Therapy: chemotherapy 65 (32.2) − radiotherapy 39 (19.3) − no therapy 98 (48.5) − The study was performed in compliance with the principles of voluntary participation and confiden- tiality, in accordance with the questioning of patients and the approval from the local Ethics Committee to study tissue samples and biological fluids. The diagnosis of lung cancer in patients has been established on the basis of clinical signs of the disease, history data, bronchoscopy, X-ray examination and computed tomography, cytomorphology of sputum and tumour tissue biopsies. In the group of NSCLC patients, the mean age was 61.6±0.6, in the control group — 61.6±0.8 years. All cases of lung malignancies have been identi- fied according to the International Classification TNM/pTNM (7th edition, 2009). The histological type of lung carcinoma was determined according to the WHO histological criteria (3rd edition, 1999). The group of NSCLC included the most common neoplasms — squamous-cell carcinoma and ade- nocarcinoma. The treatment was mostly surgical: lobectomy — 65.1%, pneumonectomy — 22.6%, lung resection — 7.0%. Only a biopsy was performed in 5.4% of cases, and in 7.9% of patients the operation was not done because of the process spread or severe comorbidity. The total DNA from the peripheral blood was isolated using the Mathew method [16]. VEGF C-634G (rs 2010963), C-2578A (rs 699947), C+936T (rs 3025039) polymorphisms were genotyped by the polymerase chain reaction method and the re- striction fragment length polymorphism analysis (PCR- RFLP analysis) using specific primers and restriction endonucleases [15, 17]. Primers for the analysis were synthesized by “Primetekh”, Minsk. The reagents for PCR and PCR-RFLP were manufactured by “Fermen- tas”, Vilnius. Statistical analysis. Statistical analyses were per- formed using Excel and Statistica 7.0. When compa- ring genotype frequencies, Pearson’s chi-squared test (χ2) was used. The association between the genotypes and the disease course was assessed by the value of odds ratio (OR). RESULTS AND DISCUSSION An elevated VEGF expression was revealed while studying the variety of malignancies: cancers of the colon, rectum, liver, lung, thyroid, digestive tract; breast, kidney and bladder adenocarcinoma; ovary and uterus carcinoma; angiosarcoma, multi- form glioblastoma [18–21]. In turn, the level of VEGF expression depends largely on the polymorphic vari- ants of the gene. Therefore, the first phase of our work was to analy- ses the association between VEGF C-634G, C-2578A and C+936T polymorphisms and the risk of NSCLC. The analysis of data on VEGF genotyping in patients with NSCLC and controls is presented in Table 2. Frequency distribution of alleles and genotypes of functionally significant VEGF polymorphisms in the population of patients living in the territory of the Republic of Belarus was close to the occur- rence of the same polymorphisms in the control group. Statistically significant differences in the frequency of the studied genotypes between the group of pa- tients with NSCLC and the control group were not 39. Schultz A, Lavie L, Hochberg I, et al. Interindividual heterogeneity in the hypoxic regulation of VEGF: significance for the development of the coronary artery collateral circulation. Circulation 1999; 100: 547–52. Experimental Oncology 37, 213–217, 2015 (September) 215 found. Similar results on the frequency of the studied VEGF single nucleotide substitutions were obtained for the populations in Germany and Sweden. Based on the results, it can be assumed that there is no connection between the studied VEGF polymor- phisms and the risk of NSCLC in this population. Since VEGF is a major mediator of angiogenesis, it can be supposed that VEGF С-634G, C-2578A and C+936T polymorphisms will rather affect the pheno- type and tumor biological behavior than the risk for tumor occurrence. The next stage of our study was to find the relation- ship between the polymorphisms under study and the clinical course of this disease. The comparative analysis of genotypes distribution of three functionally significant VEGF polymorphisms and tumor size, me- tastasis and clinical outcomes in the group of patients with NSCLC has been carried out. VEGF triggers the neoplastic angiogenesis, resulting in increased microvascular density, and malignant tis- sue receives more nutrients [23, 24]. VEGF secretion by tumor cells leads to the synthesis of proangiogenic factors. The newly formed vessels begin to supply malignant tissue with oxygen and nutrients; the tumor is growing and producing more VEGF. VEGF increases the level of VEGFR2 receptor expression by endothe- liocytes of tumor microvessels, thus stimulating the cell growth and endothelial cells proliferation [25]. Structurally and functionally the neoplastic vessels differ from the normal ones, with high permeability, chaotic branching, multiple loops, weaves, dead-end branches, lack of structured vasculature being typical for them [2]. Chaotic arrangement of tumor vessels results in uneven oxygen supply to the surrounding tissues, formation of local hypoxic foci, and is followed by tumor resistance to the radio-and chemotherapy. With an elevated VEGF expression, the vascular permeability increases leading to a higher interstitial and intratumoural pressure, facilitating the penetration of tumor cells into the bloodstream [26]. The relationship between the tumor angiogenesis level, tumor size and metastatic process is confirmed by the correlation between the course of the disease and microvascular density of the primary tumor [27– 32]. Different VEGF polymorphisms are associated with lymphogenic and hematogenous metastases in a variety of malignancies [28, 29]. The performed analysis of the relationships be- tween three VEGF gene polymorphisms and tumor size (Table 3) has shown that genotype –2578СС carriers displayed a greater degree of tumor spread (Т2–4) sig- nificantly more frequently (р = 0,002) compared to the primary focus spread (Т1). On the contrary, the carri- ers of –2578СА genotype more often presented with small non-invasive cancer (Т1). However, in this group of NSCLC patients, no statistically significant associa- tions between the studied polymorphic allele variants and regional and/or distant metastases were found (Table 4). The in vitro studies conducted by M. Moham- madi and M.С. Shahbazi has shown that -2578C allele correlated with higher VEGF expression compared to A allele [30, 31]. Similar results were obtained by M. Perrot-Applanat who proved that –2578С allele was associated with an elevated VEGF expression and pointed to the possible involvement of VEGF proteins in the autocrine regulation of the tumor growth [32]. Perhaps, this is likely to explain our findings, where the homozygous carriers (–2578СС) had higher degree of the tumor spread. The lower level of VEGF expres- sion in the presence of –2578А allele seems to have a protective effect in heterozygous carriers (–2578СА) associated with a lesser degree of the neoplasm spread. In the population under study, a significant rela- tionship between the –2578СА variant and the degree of spread of the primary tumor as well as the outcome of the disease was detected (Table 5). Heterozy- Table 2. Frequency distribution of polymorphic variants of VEGF in patients with NSCLC and control groups in different populations Genotype Belarus (original data) Germany [22] Sweden [22] Patients Controls OR (95% CI) Patients Controls OR (95% CI) Patients Controls OR (95% CI)VEGF (G634C) N=186; n (%) N=364; n (%) N, % N, % N=936; n (%) N=941; n (%) GG 83 (44.6) 186 (51.1) 0.77 (0.54–1.10) − − − 488 (52.1) 492 (52.3) 1.00 GC 88 (47.3) 154 (42.3) 1.22 (0.86–1.75) − − − 363 (38.8) 367 (39.0) 1.00 (0.82–1.21) CC 15 (8.1) 24 (6.6) 1.24 (0.64–2.43) − − − 85 (9.1) 82 (8.7) 1.05 (0.74–1.47) G allele 254 (68.3) 526 (72.3) 0.83 (0.63–1.09) − − − 1339 (71.5) 1351 (72.0) 0.99 (0.86–1.14) С allele 118 (31.8) 202 (27.7) 1.21 (0.92–1.59) − − − 533 (28.5) 531 (28.0) 1.01 (0.86–1.17) VEGF (С2578А) N=162; n (%) N=360; n (%) N=153; n (%) N=162; n (%) N=939; n (%) N=940; n (%) СС 41 (25.3) 83 (23.0) 1.13 (0.74–1.74) 44 (28.8) 50 (30.9) 1.00 258 (27.5) 257 (27.3) 1.00 СА 90 (55.6) 186 (51.7) 1.17 (0.81–1.70) 75 (49.0) 72 (44.4) 1.18 (0.68–2.06) 449 (47.8) 451 (48.0) 0.99 (0.79–1.24) АА 31 (19.1) 91 (25.3) 0.70 (0.44–1.11) 34 (22.2) 40 (24.7) 0.97 (0.50–1.86) 232 (24.7) 232 (24.7) 1.00 (0.82–1.23) C allele 172 (53.1) 352 (48.9) 1.18 (0.91–1.54) 163 (53.3) 172 (53.1) 1.01 (0.74–1.38) 965 (51.4) 965 (51.2) 0.78 (0.68–0.89) A allele 152 (46.9) 368 (51.1) 0.85 (0.65–1.10) 143 (46.7) 152 (46.9) 0.99 (0.73–1.36) 913 (48.6) 915 (48.8) 1.05 (0.93–1.20) VEGF (С936T) N=161; n (%) N=360; n (%) N=153; n (%) N=163; n (%) N=924; n (%) N=934; n (%) CC 115 (71.4) 267 (74.2) 0.87 (0.57–1.32) 120 (78.4) 128 (78.5) 1.00 708 (76.6) 720 (77.1) 1.00 CT 38 (23.6) 80 (22.2) 1.08 (0.70–1.68) 31 (20.3) 31 (19.0) 1.07 (0.59–0.93) 204 (22.1) 203 (21.7) 1.02 (0.81–1.28) TT 8 (5.0) 13 (3.6) 1.40 (0.57–3.44) 2 (1.3) 4 (2.5) 0.53 (0.07–3.46) 12 (1.3) 11 (1.2) 1.11 (0.45–2.71) С allele 268 (83.2) 614 (85.3) 0.86 (0.60–1.23) 271 (59.8) 287 (88.0) 0.20 (0.14–0.30) 1620 (87.7) 1643 (88.0) 0.97 (0.80–1.18) T allele 54 (16.8) 106 (14.7) 1.17 (0.82–1.67) 182 (40.2) 39 (12.0) 4.94 (3.37–7.25) 228 (12.3) 225 (12.0) 1.03 (0.84–1.25) Table 3. Distribution of frequencies of polymorphic variants of VEGF and tumor size in patients with NSCLC Tumour size N G634C N C2578A N C936T GG GC CC CC CA AA CC CT TT n % n % n % n % n % n % n % n % n % T1 70 32 45.7 33 47.1 5 7.1 57 6 10.5* 40 70.2** 11 19.3 41 29 70.7 11 26.8 1 2.4 216 Experimental Oncology 37, 213–217, 2015 (September) gous genotype –2578СА carriers were included into the follow-up group significantly more often (р = 0.021). It should be noted that a similar results was observed for another VEGF polymorphism — the heterozygous variant +936СT located in the 3´-UTR (р = 0.012). More- over, C. Oliveira et al. have shown that heterozygous +936СT genotype was increasingly associated with a non-recurrent disease [11]. No associations between the VEGF –634G>C polymorphism and the survival of patients with NSCLC have been found. It has been demonstrated that the polymorphism of a single nucleotide located in the promoter region or 3´- and 5´-UTR affects the protein expression at the transcriptional level [33]. Initially, the hypoxia induced factor (HIF-1) joins VEGF gene in the pro- moter region (–2578СА polymorphism) and increases its expression. Gene polymorphism in this region can weaken or strengthen this interaction, and, therefore, alter the VEGF expression (–2578СА) [34] that, in turn, will affect the formation and permeability of microves- sels and, finally, the tumor size, metastasis and survival. Based on the results of the study on the correla- tion between the polymorphisms and development of the di sease in this population, it has been estab- lished that the carriers of –2578СС genotype displayed a greater degree of tumor spread (Т2–4) significantly more often (р = 0.002) while the carriers of –2578СА genotype presented with small non-invasive can- cers (Т1) more frequently (р = 0.021). The carriers of –2578C/A genotype were significantly often observed (р = 0.021) in the “follow-up” group. Given that the high level of VEGF expression is related to an increased risk of a recurrent disease and shorter survival of subjects with different cancers [35], one may suggest the VEGF (–2578С/-) genotype to contribute to the high expression of the corresponding protein product in the development of NSCLC. The mechanism of most associations between single nucleotide substitutions and clinical behavior of tumors is still largely unclear. It should be taken into account that 5´- and 3´-UTR contain key regulatory elements sensitive to hypoxia [36–38] and contribute to high variability of VEGF production [39]. For instance, VEGF- 634G>C and –2578A>C polymorphisms in the 5´-UTR affect the efficiency of protein translation [33], and 936C>T polymorphism in the 3´-UTR influences the VEGF circulating plasma concentrations [13] and tumor tissue expression of VEGF [5]. This can explain the protective ef- fect of the VEGF +936T allele in breast cancer metastasis (due to the reduced VEGF expression) [4, 11]. Thus, the results of our study demonstrate that VEGF –2578A/C and +936C/T polymorphisms are among the factors determining the individual pecu- liarities of NSCLC course in this population and can be used for clarifying the prognosis of the disease. REFERENCES 1. Ferrara N, Gerber HP, LeCouter J. The biology of VEGF and its receptors. Nat Med 2003; 9: 669–76. 2. Ferrara N. Vascular endothelial growth factors as a target for anticancer therapy. Oncologist 2004; 9: 2–10. 3. Ferrara N. VEGF and the quest for tumour angiogenesis factors. Nat Rev Cancer 2002; 2: 795–803. 4. Jin Q, Hemminki K, Enquist K, et al. Vascular endothelial growth factor polymorphisms in relation to breast cancer deve- lopment and prognosis. Clin Cancer Res 2005; 11: 3647–53. 5. Koukourakis MI, Papazoglou D, Giatromanolaki A, et al. VEGF gene sequence variation defines VEGF gene ex- pression status and angiogenic activity in non-small cell lung cancer. Lung Cancer 2004; 46: 293–8. 6. Liu D-H, Zhang X-Y, Fan D-M, et al. Expression of vas- cular endothelial growth factor and its role in oncogenesis of hu- man gastric carcinoma. World J Gastroenterol 2001; 7: 500–5. 7. Yamamori M, Sakaeda T, Nakamura T, et al. Asso- ciation of VEGF genotype with mRNA level in colorectal adenocarcinomas. Biochem Biophys Res Commun 2004; 325: 144–50. 8. Lu H, Shu Z-O, Cui Y, et al. Association of genetic polymorphisms in the VEGF gene with breast cancer survival. Cancer Res 2005; 65: 5015–9. 9. Yang B, Cross DF, Ollerenshaw M, et al. Polymorphisms of the vascular endothelial growth factor and susceptibility to dia- betic microvascular complications in patients with type 1 diabe- tes mellitus. J Diabetes Complications 2003; 17: 1–6. 10. Langsenlehner U, Wolf G, Langsenlehner T, et al. Genetic polymorphisms in the vascular endothelial growth factor gene and breast cancer risk. The Austrian «tumour of breast tissue: incidence, genetics, and environmental risk factors» study. Breast Cancer Res Treat 2008; 109: 297–304. 11. Oliveira C, Lourenço G, Silva P, et al. Polymorphisms in the 5�- and 3�-untranslated region of the VEGF gene and sporadic breast cancer risk and clinicopathologic characte- ristics. Tumour Biol 2011; 32: 295–300. 12. Qi M, Huang X, Zhou L, Zhang J. Four polymor- phisms of VEGF (+405C>G, ‒460T>C, ‒2578C>A, and ‒1154G>A) in susceptibility to psoriasis: a meta-analysis. DNA and Cell Biology 2014; 33: 234–44. 13. Renner W, Kotschan S, Hoffmann C, et al. A common 936 C/T mitation in the gene for vascular endothelial growth factor is associated with vascular endothelial growth factor plasma levels. J Vasc Res 2000; 37: 443–8. 14. Schneider B, Radovich M, Sledge G, et al. Association of polymorphisms of angiogenesis genes with breast cancer. Breast Cancer Res Treat 2008; 111: 157–63. 15. Li Y, Liang J, Liu X, et al. Correlation of polymor- phisms of the vascular endothelial growth factor gene and the risk of lung cancer in an ethnic Han group of North China. Exp Ther Med 2012; 3: 673–6. 16. Mathew CC. The isolation of high molecular weight eucaryotic DNA. In: Methods in Molecular Biology. JMNJ Walker ed. Clifton: Human Press, 1984; 2: 31–4. 17. Lee SJ, Lee SV, Jeon HS, at al. Vascular endothelial growth factor gene polymorphisms and risk of primary lung cancer. Cancer Epidemiol Biormarkers Prev 2005; 14: 571–5. 18. Lee TH, Avraham HK, Jiang S, et al. Vascular endo- thelial growth factor modulates the transendothelial migra- tion of MDA-MB-231 breast cancer cells through regulation of brain microvascular endothelial cell permeability. J Biol Chem 2003; 278: 5277–84. 19. Ferrante M, Pierik M, Henckaerts L, et al. The role of vascular endothelial growth factor (VEGF) in inflammatory bowel disease. Inflamm Bowel Dis 2006; 12: 870–8. 20. Poon RT, Fan ST, Wong J. Clinical implications of cir- culating angiogenic factors in cancer patients. J Clin Oncol 2001; 19: 1207–25. Experimental Oncology 37, 213–217, 2015 (September) 217 21. Rosen LS. Clinical experience with angiogenesis sig- naling inhibitors: focus on vascular endothelial growth factor (VEGF) blockers. Cancer Control 2002; 9: 36–44. 22. Jin Q, Hemminki K, Enquist K, et al. Vascular en- dothelial growth factor polymorphisms in relation to breast cancer development and prognosis. Clin Cancer Res 2005; 11: 3647–53. 23. Carmeliet P, Jain R. Angiogenesis in cancer and other diseases. Nature 2000; 407: 249–57. 24. Jain RK. Normalization of tumour vascula- ture: an emerging concept in antiangiogenic therapy. Science 2005; 307: 58–62. 25. Millauer B, Shawver LK, Plate KH, et al. Glioblastoma growth inhibited in vivo by a dominant-negative Flk-1 mutant. Nature 1994; 367: 576–9. 26. Kopnin BP. Basic properties of the neoplastic cells and the underlying mechanisms of their occurrence. Pract Oncol 2002; 3: 229–35 (in Russian). 27. Folkman J. Angiogenesis. Annu Rev Med 2006; 57: 1–18. 28. Ishigami SI, Arii S, Furutani M, et al. Predictive value of vascular endothelial growth factor (VEGF) in metastasis and prognosis of human colorectal cancer. Br J Cancer 1998; 78: 1379–84. 29. Kawakami M, Furuhata T, Kimura Y, et al. Expres- sion analysis of vascular endothelial growth factors and their relationships to lymph node metastasis in human colorectal cancer. J Exp Clin Cancer Res 2003; 22: 229–37. 30. Mohammаdi M, Ollier WE, Hutchinson IV. A func- tional association study of VEGF gene promoter polymor- phisms with VEGF expression by stimulated pbm cells. Hum Immunol 2003; 64: 125. 31. Shahbazi M, Fryer AA, Pravica V, et al. Vascular endothelial growth factor gene polymorphisms are associated with acute renal allograft rejection. J Am Soc Nephrol 2002; 13: 260–4. 32. Perrot-Applanat M, Di Benedetto M. Autocrine functions of VEGF in breast tumour cells: adhesion, survival, migration and invasion. Cell Adh Migr 2012; 6: 547–53. 33. Schneider BP, Radovich M, Miller KD. The role of vas- cular endothelial growth factor genetic variability in cancer. Clin Cancer Res 2009; 15: 5297–301. 34. Sheng Z, Guo-Ping W, Cong L, Muxiang Z. Eukary- otic initiation factor 4E (eIF4E) and angiogenesis: prognostic markers for breast cancer. BMC Cancer 2006; 6: 231–43. 35. Awata T, Inoue K, Kurihara S, et al. A common polymorphism in the 50-untranslated region of the VEGF gene is associated with diabetic retinopathy in type 2 diabetes. Diabetes 2002; 51: 1635–9. 36. Watson C, Webb N, Bottomley M, Brenchley P. Identification of polymorphisms within the vascular endo- thelial growth factor (VEGF) gene: correlation with variation in VEGF protein production. Cytokine 2000; 12: 1232–5. 37. Brown LF, Detmar M, Claffey K, et al. Vascular perme- ability factor/vascular endothelial growth factor: A multifunc- tional angiogenic cytokine. In: Regulation of Angiogenesis. ID Goldberg, EM Rosen, eds. Birkhäuser Basel, 1997; 79: 233–69. 38. Minchenko A, Salceda S, Bauer T, Caro J. Hypoxia regulatory elements of the human vascular endothelial growth factor gene. Cell Mol Biol Res 1994; 40: 35–9. Copyright © Experimental Oncology, 2015