Expression Of Ang-1 And Ang-2 In Vulvar Precancer Disease And Vulvar Squamous Cell Cancinoma

ObjectiveAngiopoietin is one of the known growth factor family that is specific to vascular endothelium, including Ang-1, Ang-2, Ang-3, Ang-4. The action of angiopoietin is associated not only with angiogenesis but also with postnatal vasulogenesis. Ang-1 contributes to blood vessel maturation and stability, and Ang-2 may be involved in angiogenesis. In our research, we studied the function and expression of Ang-1 and Ang-2 in different vulvar diseases.Materials and MethodsThirty cases of vulvar squamous cell carcinoma; 35 cases of VIN (15 VINⅢ, 20VINⅠ-Ⅱ); 40 cases of NNEDV (20 of them were squamous cell hyperplasia, the other 20 were lichen sclerosus); 10 cases of normal vulvar skin. We collected the clinical materials, including age, clinical stage, pathological grades, lymph nodes metastases.In our study, immunohistochemistry was employed with SABC method. Rabbit anti-human Ang-1/-2 polyclonal antibody (from Boster Biotech) with dilution. All procedures were implemented according to the product illustration. Positive and negative controls were performed to ensure the result reliability.The immunostaining of Ang-1/-2, CD34 was localized on the cytoplasm. We counted the positive cells of five high-power microscope and averaged the number of positive cells in per high-power microscope.Statistical evaluation was performed using the Independent-Sample t Test to differentiate the number of cells among different groups. SPSS version13.0 software employed to analyze all data.Results 1. The expression of Ang-1 among different groupsAng-1 protein was localized on the cytoplasm. The number of Ang-1 protein positive cells were 0 in normal vulvar skin, 8.56±4.75 in LS, 4.67±2.43 in SH, 8.51±3.35 in VINⅠ-Ⅱ, 29.10±4.56 in VINⅢ, 38.97±5.85 in VSCC. There were differences between VINⅠ-Ⅱand VINⅢ(P＜0.05).2. The expression of Ang-2 among different groupsAng-2 protein was localized on the cytoplasm. The number of Ang-2 protein positive cells were 0 in normal vulvar skin, 7.38±3.41 in LS, 4.70±2.58 in SH, 8.23±4.43 in VINⅠ-Ⅱ, 24.66±4.79 in VINIII, 34.33±4.62 in VSCC. There were differences between VINⅠ-Ⅱand VINⅢ(P＜0.05).3. The expression of CD34 among different groupsCD34 protein was localized on cytoplasm of endothelial cells. We used MVD to count the number of positive vessels. MVD were 20.44±3.66 in normal vulvar skin; 10.34±2.83 in LS; 21.58±2.48 in SH; 21.83±3.18 in VINⅠ-Ⅱ, 34.71±4.01 in VINⅢ; 39.88±7.65 in VSCC. There were differences between normal vulvar skin and LS, VINⅢ, VSCC (P＜0.05).4. The relationship of Ang-2 with vulvar carcinoma clinical and pathological materialsThe rates of Ang-2 protein expression showed correlation with pathological grades and lymph nodes metastases (P＜0.05), but no significiant correlation with age, clinical stage.Conclusions1. Vessel form that Ang induced was at static relatively in normal vulvar skin;2. There were microcirculation abstacles in LS and it induced the expression of Ang-2 protein increased. At the same time, Ang-2 collaborated with VEGF, and they promoted vessel form;3. Ang-1 hadn't the role of vessel leakage that anti-inflammatory cytokines leaded in SH;4. Ang-1 and Ang-2 protein involved the process of vessel form in VIN;5. The expression of Ang-2 had correlated with vulvar carcinoma infiltration transfer and malignant progress.