The Tissue Microarray Assay Sparc Protein Expression And Significance In Endometrial Carcinoma

The expression of SPARC and clinical value in Endometriai CarcinomaPrefaceUnder the influence of increasing proportion of aged population,infertility less accouchement,and the wide application of exogenous estrogen etc,endometrial carcinoma has now become the most frequent gynecologic cancer in the world.The judgment of the malignant level and the metastasis potential which directly affect the biological behavior of carcinoma is essential for evaluating the prognosis of the patients.In this study,we adopted tissue array technique to detect the expression of SPARC in 86 cases of endometrial cancer tissue samples,thereafter to judge the feasibility of taking the expression of SPARC as a factor to evaluate the prognosis of patients suffering from endometrial carcinoma.Materials and Methods1.Case selectionA total of 84 paraffin-embedded tissue blocks of endometrial carcinoma were retrieved from the surgical pathology department at the affiliated of Medical College of Qingdao Univercity during 2004 to 2006.The selected clinical cases were confirmed by pathological diagnosis.All of the patients were preoperatively untreated and with no combination of other neoplasm.2.MaterialsAnti-SPARC mouse monoclonal antibody was purchased from American SANTA CRUZ Co.Ltd.3.Methods(1) Make tissue array(2) Tissue array embedded in paraffin were cut into 4m-thick sections.These sections were deparaffinized with xylene,and rehydrated with a series of graded alcohols.The slides were immersed in methanol containing 3%H₂O₂ to block endogenous peroxidase activity.After heat antigen retrieve processing,the sections were treated with normal serum to block nonspecific staining.Sections were then incubated with anti-SPARC primary antibodies respectively at 4℃overnight in a moisture chamber. This was followed by incubation with a biotinylated secondary antibody at room temperature for 35 minutes and thereafter incubation with horseradish peroxidase conjugated avidin-biotin complex at room temperature for 1 hour.Immunostaining was visualized using 3,3V-diaminobenzidine tetradrochloride substrate.Slides were counter-stainde with resin.Tissue ample of esophageal squamous cell carcinoma was used an positive control of SPARC immunoassaying respectively.PBS replacing the primary antibodies was used as negatives controls.4.Results JudgmentThe introcytoplasmic brown granule was considered as SPARC immunopositive.5.Statistical AnalysisAll the data was processed by chi-square test or Fisher exact probability test using SPSS 13.software package.ResultsThe positive rate of SPARC protein expression was 96.55%,76.79%,59.50%in normal endometrium,proliferetive endometrium and endometrial carcinoma,obviously compared with other groups,with a statistical significance(P＜0.05).There was a close relationship between the SPARC expression and the clinical and histological grades of the specimens.In endometrial carcinoma,the expression loss of SPARC protein positively was significantly associated with clinical and histological grades,the expression intensity of SPARC was related to surgical stage,histological grade,musckes invasion and lymph node metastasis(P＜0.05).ConclusionsThe expression loss of SPARC protein was well related with the generation and development of endometrial carcinoma.Detection of SPARC may provide a necessary theory basis on the prognosis and the clinical treatment of endometrial carcinoma.