Research Article - Biomedical Research (2017) Volume 28, Issue 14
Vascular endothelial growth factor (VEGF) rs3025039 polymorphism is associated with increased risk of osteosarcoma
- *Corresponding Author:
- Zongsheng Yin
Department of Orthopedics
First Affiliated Hospital of Anhui Medical University, PR China
Accepted on June 15, 2017
Background: The role of vascular endothelial growth factor (VEGF) rs3025039 polymorphism on osteosarcoma risk was not fully clear. Thus, we did a case-control study to evaluate the association between VEGF rs3025039 polymorphism and osteosarcoma risk.
Method: This study included 242 patients with osteosarcoma and 253 controls. The genotyping was conducted using the matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS).
Results: The VEGF rs3025039 TT genotype was significant higher in the osteosarcoma group than in the control group (OR=2.42, 95%CI 1.00-5.85, P=0.04). The TT genotype and CT genotype was also significantky associated with osteosarcoma risk (OR=1.43, 95%CI 1.00-2.05, P=0.04). Under the allelic model, T allele of rs3025039 was significantly associated with higher osteosarcoma risk (OR=1.41, 95%CI 1.05-1.90, P=0.02). In addition, VEGF rs3025039 TT genotype was not associated with tumor location (OR=1.02, 95%CI 0.59-1.77, P=0.94) and metastasis (OR=1.76, 95%CI 0.90-3.43, P=0.10).
Conclusion: In conclusion, this study confirmed that VEGF rs3025039 polymorphism was significantly associated with higher risk of osteosarcoma.
Vascular endothelial growth factor, Osteosarcoma, Genetic.
Osteosarcoma originates from primitive bone-forming mesenchymal cells and has been identified as an aggressive sarcoma of the bone . Osteosarcoma has a wide range of histological appearances. Conventional osteosarcoma may be classified as osteoblastic, chondroblastic or fibroblastic, depending on the predominant type of extracellular matrix present . Despite combined therapy, more than 30% of the patients showed the recurrence or metastatic disease during the first five years after diagnosis .
Vascular endothelial growth factor (VEGF) plays an important role in the maintenance of endothelial integrity, endothelial survival and the physiological function of endothelium . Niu et al. suggested that knockdown of VEGFA by siRNA inhibited proliferation, migration, and invasion of U2OS cells . Peng et al. demonstrated that VEGF silencing could suppress cells proliferation, promote cells apoptosis and reduce osteosarcoma angiogenesis through inactivation of VEGF/ PI3K/AKT signaling pathway . Han found that VEGF is related to the grade and metastasis of osteosarcoma . The human VEGF gene is located on chromosome 6p21.3 with 7 introns and 8 exons and shows polymorphism . The role of VEGF rs3025039 polymorphism on osteosarcoma risk was not fully clear. Thus, we did a case-control study to evaluate the association between VEGF rs3025039 polymorphism and osteosarcoma risk.
This study consisted of 242 patients with osteosarcoma. They were treated in the First Affiliated Hospital of Anhui Medical University, Ma’asshan hospital and Drum Tower Hospital between 2010 and 2017. The control group contained 253 healthy subjects who came from our hospital’s physical checkup center during the same period of time. All control subjects had no history of cancer. Written permission was obtained from all the participants and the study was approved by the Research Ethics Committee of First Affiliated Hospital of Anhui Medical University.
Peripheral venous blood samples (10 mL) were collected from all patients using vacutainer tubes in the morning. Blood samples (5 mL) for genetic analyses were transferred into tubes which contained ethylenediamine tetra-acetic acid (EDTA). Genomic DNAs were isolated using genomic DNA extraction kit (QIA amp DNA Blood Mini Kit, Qiagen, Berlin, Germany) under the manufacturer's instructions.
The SNP genotyping was conducted using the matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) (Sequenom Inc., San Diego, CA, USA). The PCR reaction was performed in a 20 μl reaction mixture containing 40-160 ng DNA template, 0.25 units Taq DNA polymerase (Sangon Biotech), 250 μM each deoxyribonucleotide triphosphate (dNTP) (Sangon Biotech), 0.5 μM forward primer, 0.5 μM reverse primer, and 1X PCR buffer with 1.2 mM MgCl2. The SNaPshot assay (Applied Biosystems) was performed to confirm genotypes of 6 DNA samples. All genotyping procedures were carried out in a double-blind manner and the whole assays were proved to be reliable.
Data are presented as mean ± standard deviation (SD) or number (percentage). The chi-square test and Student’s t-test were used to compare case and control groups, as appropriate. The statistical comparison between 2 groups was conducted using the t-test or the analysis of variance (ANOVA). Genotype distribution in the control group was tested by Hardy-Weinberg equilibrium (HWE). The differences in genotype and allele distribution between the patients and the controls are represented as odds ratio (OR) and 95% confidence interval (CI). P values for all tests are 2-tailed, and <0.05 was considered as statistically significant. Statistical analysis was conducted using SPSS 18 software (SPSS Inc., Chicago, IL, USA).
Table 1 shows the demographic and clinical characteristics of 242 patients with osteosarcoma and 253 controls. Comparisons between the osteosarcoma group and the control group demonstrated that family history of cancer was significantly higher in patients with osteosarcoma than in controls (P<0.05). No statistical differences were seen in age and gender (all P>0.05).
The genotype and allele frequencies of the VEGF rs3025039 polymorphism in patients and controls are displayed in Table 2. The VEGF rs3025039 TT genotype was significant higher in the osteosarcoma group than in the control group (OR=2.42, 95% CI 1.00-5.85, P=0.04). The TT genotype and CT genotype was also significantly associated with osteosarcoma risk (OR=1.43, 95% CI 1.00-2.05, P=0.04). Under the allelic model, T allele of rs3025039 was significantly associated with higher osteosarcoma risk (OR=1.41, 95% CI 1.05-1.90, P=0.02). In addition, VEGF rs3025039 TT genotype was not associated with tumor location (OR=1.02, 95% CI 0.59-1.77, P=0.94; Table 3) and metastasis (OR=1.76, 95% CI 0.90-3.43, P=0.10; Table 3).
|Characteristics||Case (n=242)||Control (n=253)||P value|
|Age at diagnosis||26.1 ± 16.6||28.1 ± 12.2||0.17|
|Family history of cancer||<0.01|
Table 1. Characteristics of the cases and controls.
|Genotype||Case||Control||OR||95% CI||P value|
Table 2. The genotype and allele frequencies of VEGF polymorphism in the cases and controls.
|Characteristics||TT+CT (n=111)||CC (n=94)||OR||95% CI||P value|
Table 3. Relation of VEGF polymorphism and characteristics.
In the current study, we investigated the association between the VEGF rs3025039 polymorphism and osteosarcoma risk. The VEGF rs3025039 TT genotype was significant higher in the osteosarcoma group than in the control group. The TT genotype and CT genotype was also significantky associated with osteosarcoma risk. Under the allelic model, T allele of rs3025039 was significantly associated with higher osteosarcoma risk. In addition, VEGF rs3025039 TT genotype was not associated with tumor location and metastasis.
Zhang et al. found that vascular endothelial growth factor overexpression predicted a worse prognosis for laryngeal cancer patients . Zhao et al. indicated that high plasma/ intratumoral VEGF-A level at baseline could predict poor treatment effect (depressed PFS and OS) of BEV-based chemotherapy in mCRC . Cai et al. found that high level of VEGF is associated with poor outcomes in HCC patients treated with sorafenib . Hui et al. indicated that the OS of the VEGF-positive group with ovarian cancer was significantly poorer than the VEGF-negative group . Ma et al. suggested that both the VEGF +936C/T and -634G/C polymorphisms influence breast cancer susceptibility and tumor growth, instead of metastasis .
This study has some limitations. The selective bias was mostly controlled by the design of a hospital-based case-control study. As other case-control studies, this study raises concern about recall bias and residual confounding. The major difficult is still the inability to separate exposures to factors prior to clinical onset from exposures to factors after clinical onset.
In conclusion, this study confirmed that VEGF rs3025039 polymorphism was significantly associated with higher risk of osteosarcoma risk.
- Maire G, Martin JW, Yoshimoto M, Chilton-MacNeill S, Zielenska M, Squire JA. Analysis of miRNA-gene expression-genomic profiles reveals complex mechanisms of microRNA deregulation in osteosarcoma. Cancer Genet 2011; 204: 138-146.
- Marina N, Gebhardt M, Teot L, Gorlick R. Biology and therapeutic advances for pediatric osteosarcoma. Oncologist 2004; 9: 422-441.
- Ottaviani G, Jaffe N. The epidemiology of osteosarcoma. Cancer Treat Res 2009; 152: 3-13.
- Ferrara N. Molecular and biological properties of vascular endothelial growth factor. J Mol Med (Berl) 1999; 77: 527-543.
- Niu J, Sun Y, Guo Q, Niu D, Liu B. miR-1 Inhibits Cell Growth, Migration, and Invasion by Targeting VEGFA in Osteosarcoma Cells. Dis Markers 2016; 2016: 7068986.
- Peng N, Gao S, Guo X, Wang G, Cheng C, Li M, Liu K. Silencing of VEGF inhibits human osteosarcoma angiogenesis and promotes cell apoptosis via VEGF/PI3K/AKT signaling pathway. Am J Transl Res 2016; 8: 1005-1015.
- Han G, Wang Y, Bi W, Jia J, Wang W, Xu M. Effects of vascular endothelial growth factor expression on pathological characteristics and prognosis of osteosarcoma. Clin Exp Med 2016; 16: 577-584.
- Arcondéguy T, Lacazette E, Millevoi S, Prats H, Touriol C. VEGF-A mRNA processing, stability and translation: a paradigm for intricate regulation of gene expression at the post-transcriptional level. Nucleic Acids Res 2013; 41: 7997-8010.
- Zhang LP, Chen HL. Increased vascular endothelial growth factor expression predicts a worse prognosis for laryngeal cancer patients: a meta-analysis. J Laryngol Otol 2017; 131: 44-50.
- Zhao L, Zhang D, Ma H, Jin M, Huang F, Zhang T. High VEGF-A level at baseline predicts poor treatment effect of bevacizumab-based chemotherapy in metastatic colorectal cancer: a meta-analysis. Panminerva Med 2016; 58: 48-58.
- Cao G, Li X, Qin C, Li J. Prognostic Value of VEGF in Hepatocellular Carcinoma Patients Treated with Sorafenib: A Meta-Analysis. Med Sci Monit 2015; 21: 3144-3151.
- Hui G, Meng M. Prognostic value of vascular endothelial growth factor expression in women with ovarian cancer: A meta-analysis. J BUON 2015; 20: 870-878.
- Ma J, Hu W, Zhang P, Sun Y, Wang N, Teng X, Qiao Q. The Association Between VEGF +936C/T and -634G/C Polymorphisms and Breast Cancer Susceptibility, Tumor Growth, and Metastases: Evidence From 20,728 Subjects. Cancer Invest 2015; 33: 312-317.