Expression And Signifiance Of Ki67 And Caspase-3 In Vulvar Neoplasia

PrefaceThe proliferation and apoptosis of cell aire correlated with tumor genesis, invasion and metastasis. An imbalance of cell apoptosis and proliferation was found during the multistep and multifactorial process. It is crucial to estimate the cell proliferation and apoptosis for study of behavior of tumor. Ki67 is a new cell cycle related to cell proliferation nuclear antigen and has been employed as a po-tiential marker of cell proliferation. Expression of Ki67 is in the growth and synthesis phases of the cell cycle, namely, G₁, S, G₂, and M phase, but not in resting G₀ phase. Caspases are the key executioners of apoptosis. It has been repor-tad that Caspase - 3 , a cell death protease, is most directly correlated with apoptosis because of its central location in the proteases cascade pathway , and can hydrolyze numerous of protein substrates involving in cell biological and mor-pholigic functions, which directly leads to apoptosis. It was aim of this study to investigate the expression of clinical significance of proliferating cell nuclear antigen Ki67 and protein related cell apoptosis Caspase - 3 staining in normol vulvar epithelia , nonneoplastic epithelial disorders of vulvar (NNED) , vulvar in-traepithelial neoplasia ( VIN) , as well as squamous cell carcinomas of the vulvar (VSCC).Material and MethodsThirty cases of vulval squamous carcinoma;30 cases of VIN(10 VIN I ,10 VIN II ,10 VIN III) ;30 cases of NNED, 15 of them were squamous cell hyper-plasia, the other 15 were lichen sclerosus;10 cases of normal vulvar skin. We collected the clinical material: age , clinical stage , histologic grade , nodal status.In our study , immunohistochemistry was employed with SP method. Mouse anti — human Ki67 monoclonal antibody ( from Maixim Biotech) was applied without dilution, whereas rabbit anti - human Caspase - 3 polyclonal antibody ( from NeoMarker Biotech) were diluted in phosphate - buffered saline at dilution ratio of 1:200. All procedures were implemented according to the product illustration . Positive and negative controls were performed to ensure the results' reliability.The immunostaining of Ki67 was localized on the nuclei and caspase - 3 was on cytoplasm or nuclei. A combined score for the immunostaining, based on both qualitative and quantitative immunostaining ,was composed by adding both the qualitative and quantitative score which was then divided into three group ; score 0—2 ( - ) ; score3 ( + ) ; score > 3 ( + + ).Statistical evaluation was performed using chi — square to differentiate the rates of different groups, SPSS version 15.0 software was employed to analyze all data,ResultsNonsignificant difference was found between the exprssion of Ki67 in nor-mol vulvar epithelia and lichen sclerosus of vulvar (10% vs6.7%)(P>0.05). There was nonsignificant difference among VIN I - II and squamous cell hyper-plasia(50% vs 46. 7% ) ( P > 0. 05 ) , but there was a significant difference between VIN I II and VIN III (50% vs 90% ) ( P <0.05). There was nonsignificant difference between the exprssion of Ki67 in VSCC and VIN III (83. 3% vs 90% ) ( P > 0.05). There was a significant difference between VIN I II and normol vulvar epithelia (50% vs 10% ) ( P < 0. 05 ). The expression of Ki67in VSCC was not related with age ,clinical stage, lymph node metastases or the pathological grade( P > 0.05).Caspase - 3 protein positive rate was 53. 3% in vulvar squamous carcino-...