Study On The Association Of VEGF And Its Single Nucleotide Polymorphisms With The Risk Of Developing Endometriosis

ObjectiveThe purpose of this experiment is to detect the expression of serum vascular endothelial growth factor in patients with endometriosis, discussing the role of VEGF in the development of endometriosis and its clinical significance preliminarily; detect whether the single nucleotide polymorphisms of VEGF promoter region -2578C/A,-1154G/A,-460C/T and +936C/T are existed in the local crowds, analyzing if there was a difference about the following gene loci genotypes distribution between patients with endometriosis and the controls, in the purpose of further exploring the relationship between polymorphisms of VEGF promoter region and the risk of developing endometriosis; laying the theoretical foundation of further study and the treatment aiming at VEGF antagonist.MethodsThe method of case-control study is adopted by this experiment. We collected serum samples through thirty-six patients with endometriosis randomly, thirty-six control samples in the corresponding period, and twenty-two serum samples through patients with the treatment of gestrinone after surgery, the expression levels of serum VEGF was detected by enzyme linked immunosorbent assay, in the purpose of comparing the difference between patients with different clinical symptoms and stages, we also compared the levels of serum VEGF with patients before or after the treatment. The genomic DNA was extracted by using proteinase K digestion followed by a salting out procedure, genotypes of VEGF -2578C/A,-1154G/A,-460C/T and +936C/T genes were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.Results1.Compared with the control group, endometriosis group had no significant difference about agesensitivity(P>0.05),but there were significant differences between the two groups about the symptoms of infertility and dysmenorrhea(P<0.05).The distribution of the VEGF -2578C/A,-1154G/A,-460C/T and +936C/T genotypes in the control group didn't deviate from that expected for Hardy-Weinberg equilibrium significantly(P>0.05).2.Comparison with the level of serum VEGF: The level of serum VEGF in endometriosis group was significantly higher than that in control group (p<0.05),the patients in endometriosis group with severe dysmenorrhea had high level of serum VEGF(p<0.05), the level of serum VEGF of patients with late clinical stages was significantly higher than that of patients with early clinical stages,the level of serum VEGF of patients were obviously higher than that after the treatment of surgery.3.In the endometriosis group, the C allele frequencies was 81.9% of VEGF -2578C/A genotype, significantly higher than the control group(73.6%);the genotype frequencies of the VEGF C/C,C/A and A/A in endometriosis and control group were 66.7%,30.6%,2.7% and 55.6%,36.1%,8.3% respectively, there were significant differences in the genotype and allele distributions between two groups(P<0.05); compared with the C/A+A/A genotypes, the C/C genotype could significantly increased the risk of developing endometriosis(OR=1.45,95%CI:1.08～2.00).4.In the endometriosis group, the G allele frequencies was 83.3% of VEGF -1154G/A genotype, significantly higher than the control group(77.8%);the genotype frequencies of the VEGF G/G,G/A and A/A in endometriosis and control group were 69.4%,27.8%,2.8% and 69.4%,27.8%,2.8% respectively, there were significant differences in the genotype and allele distributions between two groups(P<0.05); compared with the G/A+A/A genotypes, the G/G genotype could significantly increased the risk of developing endometriosis(OR=1.41,95%CI:1.03～1.95).5. The C allele frequencies was 76.4% and 79.2% in the endometriosis and control group of VEGF -460C/T genotype, the genotype frequencies of the C/C,C/T,T/T were 58.3%,36.1%,5.6% and 63.9%,30.6%,5.6% respectively, there were no statistically significant differences in the genotype and allele distributions between two groups(P>0.05); compared with the C/T+T/T genotypes, the T/T genotype could not significantly modify the risk of developing endometriosis(OR=0.86,95%CI:0.55～1.06).6. The C allele frequencies was 81.9% and 83.3% in the endometriosis and control group of VEGF +936C/T genotype, the genotype frequencies of the C/C,C/T,T/T were 66.7%,30.6%,2.8% and 69.4%,27.8%,2.8% respectively, there were no statistically significant differences in the genotype and allele distributions between two groups(P>0.05); compared with the C/T+T/T genotypes, the C/C genotype could not significantly modify the risk of developing endometriosis(OR=0.93,95%CI:0.75～1.33).Conclusions1.The expression level of serum VEGF may be associated with the endometriosis clinical stages and dysmenorrhea degree, prompting that angiogenesis is a very important factor for the development of endometriosis,monitoring the concentration of serum VEGF could become a non-invasive auxiliary method for diagnosing endometriosis.2.The level of serum VEGF may be associated with prognosis of the disease, monitoring the variation of serum VEGF concentration could be an important method for observing effectiveness and evaluating recurrence, aiming at the treatment of VEGF antagonist could become extremely the new effective treatment strategies on the basis of existing drugs and operations.3.The VEGF promoter region -2578C/A,-1154G/A,-460C/T and +936C/T single nucleotide polymorphisms were universally existed in local crowd.4.The VEGF promoter region -2578C/A and -1154G/A single nucleotide polymorphisms could be associated with the risk of developing endometriosis, the C/C and G/G genotypes could significantly increase the risk of developing endometriosis.5. No significant associations of the VEGF promoter region -460C/T and +936C/T single nucleotide polymorphisms with the risk of developing endometirosis.