Expression And Significance Of Inducible And Endothelial Nitric Oxide Synthase In Vulvar Precancer Disease And Vulvar Squamous Cell Carcinoma

ObjectiveNO has the ruction in angiogenesis by various routes. NOS, as the key enzyme forsynthesizing NO, has a cerrelation in vasodilation, angiogenesis and neoplasia. NOSincludes three isoenzymes, iNOS, eNOS and nNOS. CD34 is a sensitive endothelialmarker. Staining for CD34 has been used to measure MVD in tissues. The aim of thisthesis was to study the expression of iNOS and eNOS in VSCC, VIN, NNEDV andnormal skin in order to find their relation.Materials and MethodsMaterials: Tissue specimens from patients who had VSCC, VIN or NNEDV atthe NO.1 affiliated hospital of China Medical University from 1996 to 2006 wereretrospective examined, including 30 VSCC, 35 VIN (15 VINⅢ, 20 VINⅠ～Ⅱ), 30SH, 30 LS and 10 normal vulvar skin as control group. We collected the clinicalmaterials: age, clinical stage, histologic grade and nodal status.Main agents: Rabbit anti-human iNOS, eNOS polyclonal antibody (workingconcentration 1:100), rabbit anti-human CD34 polyclonal antibody (workingconcentration 1:80) and SABC agent box.Method: Immunohistochemistry SABC method.Result judgement: The immunostaining of iNOS, eNOS and CD34 was localizedon the cytoplasm. By DAB color, it is positive if the cytoplasma is brown. Counts were carried out in a field measuring at a magnification of 40×10 in order to obtain exactquantification of iNOS-positive cells, eNOS-positive cells and MVD.Statistical method: Statistical analyses of protein expression was carried out theIndependent-Sample T Test and Kappa Test by SPSS 13.0. Differences were consideredsignificant when P values were less than 0.05.Results1. The expression of iNOS among different groupsiNOS protein positive cells were 0 in normal vulvar skin, 8.00±3.35 in LS, 14.83±3.79 in SH, 13.60±4.16 in VINⅠ～Ⅱ, 20.13±4.91 in VINⅢand 44.18±5.08 inVSCC. There were differences between SH and LS (P＜0.05), and differences betweenthe two groups ofVIN (P＜0.05).2. The expression of eNOS among different groupseNOS protein positive cells were 0 in normal vulvar skin, 6.43±3.87 in LS, 17.86±4.82 in SH, 20.57±4.72 inVINⅠ～Ⅱ, 21.75±5.28 inVINⅢand 20.82±3.43 inVSCC. There were differences between SH and LS (P＜0.05), but no significantdifferences between the two groups of VIN (P＞0.05).3. The expression of CD34 among different groups and MVDThe MVD were 20.44±3.66 in normal vulvar skin, 10.34±2.83 in LS, 21.58±2.48 in SH, 31.91±3.93 in VIN (21.83±3.18 in VINⅠ～Ⅱ, 34.71±4.01 in VINⅢ)and 39.88±7.65 in VSCC. There were significant differences between SH and LS (P＜0.05), differences between LS and normal vulvar skin (P＜0.05), differences betweenthe two groups ofVIN (P＜0.05) and differences between VSCC and VIN (P＜0.05).4. The relationship of iNOS and eNOS with VSCC clinical andpathological materialsThe expression of iNOS showed significant correlation with clinial stages, pathol-ogical grades and lymph nodes metastases (P＜0.05). The expression of eNOS showed no significant correlation with age, clinical stages, pathological grades and lymph node-s metastases (P＞0.05).5.The relationship of iNOS and eNOS with MVD in VSCCThe VSCC MVD of iNOS between positive cells (46.35±4.36) and negative cells(31.50±4.77), there were differences (P＜0.05). But in eNOS, there were no differenc-es between positive cells (42.25±6.50) and negative cells (38.56±9.67) (P＞0.05).The VSCC MVD were positively correlated with the expression of iNOS, but not ofeNOS.6. The relationship between iNOS and eNOS in NNEDVIn NNEDV, iNOS and eNOS were both positive for 17 cases, iNOS positive butnegative for eNOS was 1 case, eNOS positive but negative for iNOS were 4 cases. Theexpression of iNOS and eNOS showed significant correlation (Kappa=0.811, P＜0.05).Conclusions1. INOS and eNOS may participated the inflammatory process ofvasodilation inSH's dermal.2. INOS and eNOS may have the fuction of vascular dilation in LS, so they couldimprove the blood transport of microcirculation compensatively.3. INOS may participate the angiogenesis process in VIN's dermal.4. INOS may have a positive correlation in angiogenesis of VSCC. The expressionof iNOS showed significant correlation with clinial stages, pathological grades andlymph nodes metastases. So iNOS may be correlated with the malignant progression ofVSCC.