Expressions Of P27, CyclinE And PCNA And Their Clinical Significance In Epithelial Ovarian Tumor

Objective: Ovarian tumor is a common disease in women reproductive system. Moreover, malignant ovarian tumor is also one of the three types of women malignant tumor. Lacking of effective diagnosis and therapy today, malignant ovarian tumor has a poor 5-year survival rate, about 25-30%. With the development of diagnosis technology and the improvement of treatment in the carcinoma of cervix and endometrium, ovarian carcinoma has been one of the most critical tumors to endanger women. It is difficult to prevent and treat ovarian cancer because its mechanism of occurrence and development is unclear. So it is vital important to understand its occurrence and development for the diagnosis, treatment and prognosis. The issue of cell cycle regulation and control is hot topic in Oncology at present. Its molecular mechanism is that the cell cycle is controlled by interaction of cyclins, cyclin-dependent kinases and CDKIs, which play an elemental role in cell cycle progress. At present, it is regarded that p27and CyclinE play an important role in the occurrence and development of ovarian cancer. PCNA is one of the best known endogenous proliferation markers. PCNA expression appeared to moredirectly correlate with the presence of proliferating cells. In the mechanism of cell cycle regulation and control, the research of p27 and CyclinE expressions has been a hot topic in epithelial ovarian carcinoma. In this study we examined the expressions of p27, CyclinE and PCNA in benign, borderline and malignant ovarian tumor with the technology of Flow Cytometry and Immunofluorescence. We also analyzed the relationship between their expressions in epithelial ovarian tumor and clinical stage, tumor grade, histological type. Meantime we analyzed cellular and subcellular localization of p27, cyclinE and PCNA expressions with the immunohistochemistry. The aim of the study is to further explore the role of p27, cyclinE and PCNA expressions and correlation with ovarian carcinoma in occurrence and development of epithelial ovarian carcinoma, and to provide theoretical basic for monitoring therapy and prognosis assess.Methods: Sixty-four patients who had been in hospital were selected from January 2002 to October 2003, age range from 34 to 70 (mean age 58.4) , of these cases 10 cases benign tumor, 12 cases borderline tumor and 42 cases malignant tumor (15 cases mucinous tumor, 14 cases serous tumor, 13 cases endometriod tumor). With FIGO staging there are four stages, including 7 cases I stage, 9 cases II stage, 23 cases IH stage, 3 cases IV stage, and 10 cases normal tissue as a control group. The technology of Flow cytometry immunoflurescence and immunohistochemistry was used. (1) Fixed ovarian tumorand normal ovarian tissue were executed single cell suspension(l X 106 cells/ml). Cell suspension was centrifuged (5min, lOOOr/min) and wished twice with 0.9% NaCl solution. Samples from 64 cases were selected for each antibody labeling (Monoclonal Mouse Anti-p27kipl protein, Monoclonal Mouse Anti-CyclinE and Monoclonal Mouse Anti-proliferating cell nuclear Antigen; working concentration 1:100). Each sample was mixed up and incubated in water-bath for 30-min at 37°C. The samples were then washed twice with PBS and incubated in water -bath for 30 min at 37°C with lOOul of second antibody of FlTC-conjugated goat anti-mouse/rabbit IgG. The cell suspension was washed and resuspended in 1.0ml PBS, filtered through nylon mesh, analyzed by flow cytometry, before the samples were analyzed. (2) One sample was fixed in natural-buffered 4% Formalin and embedded in paraffin and then diagnosed by the pathologist. Then the samples were pursued S-P immunohistochemistry staining to identity expression of P27, CyclinE and PCNA.Results: 1 Expressions of p27, CyclinE and PCNA in different ovarian tissue : Expression of p27 is low (38.9%) in epithelial ovarian carcinoma, while high in benign (80%) or borderline (75%) epithelial ovarian tumor. FI value of ovarian carcinoma (0.876 + 0.063) is obviously lower than that of benign (0.978 ± 0.009 ) and borderline (0.967 ± 0.038) ovarian tumor PO.01. Expression of CyclinE is high (76.2%) in epithelial ovarian carcinoma, while low in benign(41.7%) or borderline (30%) epithelial ovarian tumor. FI value of ovarian carcinoma (1.325 ±0.095) is obviously higher than that of benign (0.978 + 0.009) and borderline(1.012+0.042) ovarian tumor P<0.01. Expression of PCNA is high (88.1%) in epithelial ovarian carcinoma, while low in benign (10%) or borderline (33.3%) epithelial ovarian tumor. FI value of ovarian carcinoma (1.204 + 0.130) is obviously higher than that of benign (1.045+0.035) ) and borderline ( 1.105 + 0.056 ) ovarian tumor P<0.05. 2 Expressions of p27, CyclinE and PCNA in different clinical stage, tumor grade and histological type: Expression of p27 is lower in ITT-IV stage (0.851+0.056) than that of in I -II stage (0.941+0.032) ovarian carcinoma P<0.0. Expressions of CyclinE and PCNA are higher in III-IVstage (1.339 + 0.051,1.258 + 0.083) than that of in I - II stage (1.230 + 0.068,1.101 + 0.086 ) ovarian carcinoma PO.05, P<0.01. Expression of p27 is positively related to differentiation degree while expressions of CyclinE and PCNA are negative. There is no relation between expressions of p27, CyclinE and PCNA and patho-histological type. 3 The correlation of p27 and CyclinE expressions with PCNA expression in ovarian carcinoma: Expression of p27 is negatively correlated with CyclinE and PCNA r=-0.564 PO.01, r=-0.532 PO.01. The correlation of PCNA and CyclinE expressions is positive r=0.556 P<0.05. 4 Immunohistochemical localization of p27, CyclinE and PCNA in ovarian carcinoma: The subcellular localization of p27 and...