The Expression Of VEGF, HIF-1α, MMP-9 And Their Correlations With Angiogenesis In Condyloma Acuminatum

Condyloma acuminatum(CA) is a common sexually transmitted disease caused by human papilloma virus (HPV).The incidence of CA is the second one in sexually transmitted disease and increasing gradually in recently years. HPV, a kind of DNA virus, ususally infect skin, genital tract, respiratory tract and epithelial tissues of digestive and have correlation with many kinds of benign and malignant human tumors. The mainly clinical manifestation of CA is cauliflower or papilloma neoplasm in external genital organs. CA is harm to not only physical and mental health of patients with CA but also the stabilization of society and family because of its strong infectivity, high recurrence rate, and possible of ability to induce cancer.Recently, many reports reveal that angiogenesis is necessary for the growth and metastasis of all solid tumors. The volume of tumor is less than 1-2mm~3, if nutrition and oxygen supply only depend on diffuse without angiogenesis. Therefore, it is important to study the role of angiogenesis in CA because CA ,as a benign tumor, is related to angiogenesis. MVD (Microvessel density, MVD ) can reflect the status of tumor angiogenesis, CD34 is a specificness matker of vascular endothelial cell.Cell cytocin play a key role on angiogenesis. Vascular endothelial growth factor (VEGF) is the important one, a most typical mitogen of proliferation of endothelial cells, promotes angiogenesis in tumor. HIF-1 (hypoxia-inducible factor-1, HIF-1) is a central regulating factor generated from blood vessel when the tumor is hypoxia, which takes part in the total course of angiogenesis directly by affecting the expression of other vascular growth factors. The degrading of ECM (extracellular matrix,ECM) is a key step in angiogenesis . MMP (matrix metalloprotemase, MMP) is an important group of zinc depended incision enzyme which could degrade ECM. MMP-9 is the outstanding one in the course of metabolism of ECM and the angiogenesis.Many reports about the effects of HIF-1αand MMP-9 on tumor were seen, but rare reports about the relationship of MVD and HIF-1αor HIF-11αin CA. The relationship of VEGF, HIF-1αand MMP-9 is also unclear now. In order to explore the expression of VEGF, HIF-1αand MMP-9 in CA and the relationships between the above factors and angiogenesis of CA, the expression of VEGF, HIF-1α, MMP-9 and MVD in CA were detected by the immunohistochemistry, respectively.Materials and Methods1 Materials: Specimens of CA were obtained from 35 outpatients in the department of dermatology and venereology of the first affiliated hospital, Zhengzhou University. There were 23 males and 12 females in the group of CA, ages from 19 years old to 62 years old ( mean 30.8 years). The course ranges from 20 days to 140 days, mean 52.6 days. All cases of the clinical and laboratory diagnosis were made by typical clinic manifestation, confirmed morbid indication, and evidence of HPV infection. The specimens of control group derived from the normal foreskin tissues of 15 adult males who underwent surgery.2 Methods: The monoclonal antibodies were used to detect the expression of HIF-1α, HIF-1αand VEGF protein, and anti-CD34 monoclonal antibodies were used to labeled MVD. The cases were stained with immunohistochemistry SP method. According to stain level and positive cells, the expression of HIF-1α, MMP-9, VEGF were divided into four ranks: negative (-), weak positive(+):3 points, positive(++):4-5 points, strong positive(+++):6-7 points. MVD is indicated by the mean numbers of vessels of 5 high power fields.3 Statistical analysis: Statistical analysis was executed by SPSS11.0 statistical package. The categorical variable was analyzed by Chi-square and spearman rank correlation analysis. The numerical variable was analyzed by T-test and spearmanrank correlation analysis. P value of less than 0.05 was considered significant.Results1. The expression of CD34 in CA tissues and normal foreskin tissues.Staining of CD34 was easy to identify because of its high specificity, sensitivity and clear background. MVD in CA tissues and normal skin tissues was 42.52±5.60, and 26.36±6.60, respectively. There was also significantly difference between two groups(t=13.361, P<0.01).2. The expression of VEGF protein in CA tissues and normal foreskin tissues.All of the positive rate of VEGF in CA tissues and control group were 100% (35/35, 15/15). There was no significant difference between two groups (P>0.05). The distribution of VEGF in normal foreskin tissues was mainly in the basal layer, while that in CA throughout the epidermis expect the straturn corneum. The intensity of VEGF expression in CA was significant higher (++-+++) than that in normal controls (+-++) (x~2=17.388,P < 0.01).3. The expression of HIF-1αprotein in CA tissues and normal foreskin tissues.The positive rate of HIF-1αin the 35 cases of CA tissues was 94.3%(33/35), while that in normal control 66.7%(10/15). There was a significant difference between two groups(x~2 =4.556,P < 0.05). The intensity of HIF-1αexpression in CA tissues was significantly higher (++-+++) than that in normal controls (+-++) (x~2 =20.291,P <0.01)4. The expression of MMP-9 protein in CA tissues and normal foreskin tissues.The positive rate of MMP-9 in CA tissues was 100%(35/35), while that in normal control 86.7%(13/15). There was no significant difference between two groups(x~2 =2.009,P> 0.05). The intensity of MMP-9 expression in CA tissues was significantly higher (++-+++) than that in normal controls (+-++) (x~2=21.805,P <0.01).5. The correlation analyse of expression of VEGF and MVD in the CA tissues.There existed positive correlation between VEGF expression and MVD, HIF-1αexpression and MVD, and MMP-9 expression and MVD in CA tissues. Moreover, the value of MVD increased with the intensity of VEGF or HIF-1αor MMP-9 expression in the CA tissues. (r=0. 895,0.748,0.735,P<0.01).6. The correlation analyse of expression of VEGF, HIF-1αand MMP-9 in the CA tissues.Positive correlation was observed between the expression of VEGF and HIF-1α, the expression of VEGF and MMP-9, and the expression of HIF-1αand MMP-9 in CA tissues, respectively(r=0.828,0.748,0.664,P<0.01).Conclusions1. The level of VEGF, HIF-1α, MMP-9 expression are significantly higher in CA tissues than that in the control group, All of VEGF, HIF-1α, MMP-9 may play a role in the development of CA.2. There exist positive correlation between VEGF expression and MVD, HIF-1αexpression and MVD, and MMP-9 expression and MVD in CA tissues, which suggests that VEGF, HIF-1αand MMP-9 may promote angiogenesis and lead to the growth of CA induce.3. Positive correlation between VEGF and HIF-1αexpression in CA tissue shows HIF-1αin CA tissue may be the important reason of high CA endothelial cells and suggests that HIF-1αcould activate the transcription of VEGF in CA tissue, and up-regulate the expression of VEGF, then participate in CA angiogenesis.4. Positive correlation between MMP-9 and HIF-1αor VEGF expression in CA tissue suggests that MMP-9 could co-promote the growth and angiogenesis in CAtissues.