Expression Of APR3, NFκB/p65and NFAT3in Ovarian Cancer And Its Correlation Analysis

Objective:The purpose of this research is to study：1.Study on the apoptosis relatedprotein3(APR3), nuclear factor kappa B/p65(NFκB/p65), nuclear factor ofactivated T3(NFAT3) expression in epithelial ovarian tumor and itscorrelation analysis.To explore its relationship with the clinical andpathological features of ovarian epithelial carcinoma, clarify its place in thedevelopment and prognosis role of ovarian cancer.2. To explore itsrelationship with the clinical and pathological features of ovarian epithelialcarcinoma, clarify its place in the development and prognosis role of ovariancancer.3.Preliminary validation of ovarian cancer APR3existed and theexpression of APR3and P65，NFAT3in the occurrence mechanism ofovarian cancer, in order to obtain the APR3gene and pathological evidence ofovarian cancer, clinicalresearch is helpful to the further study of APR3gene,molecular treatment forovarian cancer provides a new train of thought.Materials and Methods:1.Research APR3, NFκB/p65, NFAT3protein in60cases of ovariancancer,18cases of borderline ovarian tumors and37cases of benign ovariantumors expression using immunohistochemistry.2.With different concentrations of cisplatin in human ovarian cancer celllines were SKOV3、3AO，MTT were detected using different concentrationsof cisplatin on human ovarian cancer cell line SKOV3,3AO proliferationinhibition rate. 3.The protain’s performance of the APR3, NFκB/p65, NFAT3in humanovarian cancer cell line SKOV-3,3AO by before and after usingimmunocytochemistry cisplatin.4.The protain’s performance of the APR3, NFκB/p65, NFAT3in humanovarian cancer cell line SKOV3,3AO by before and after using western blotmethod to detect cisplatin.Results:1.The protain’s performance of the APR3, NFκB/p65, NFAT3in humanovarian cancer cell:(1) The positive expression rate APR3protein in ovariancarcinoma70.0%, significantly higher than borderline ovarian tumors (55.6%)and benign epithelial ovarian (43.2%), there was significant difference (P<0.05); The expression of APR3and tissue differentiation, clinical stage,lymph node metastasis and ascites, abdominal viscera and the positiveexpression rate of APR3with decreasing ovarian cancer tissue differentiationand significantly increased (P<0.05), The later the clinical staging of thehigher expression (P<0.05), Abdominal organs and lymph node metastasispositive rate than those without abdominal organs and lymph node metastasiswere higher (P<0.05), Ascites than those without ascites were higher (P<0.05).(2) the positive expression of NF κ B/p65protein in ovarian cancer tissue was61.7%, significantly higher than that of borderline ovarian tumor (38.9%) andbenign ovarian epithelial tumor (29.7%), there was significant difference(P<0.05), NF κ B/p65expression and tissue differentiation, clinical stage, theabdominal organs and lymph node metastasis and ascites: lower histological,its expression is higher (P<0.05); clinical staging of advanced rate higher thanthe early positive expression (P<0.05). The positive expression of abdominalorgans and lymph node metastasis were higher than those without metastasis (P<0.05); the positive expression rate than those without ascites ascites(P<0.05).(3) the positive expression of NFAT3protein in ovarian cancertissue was50%, significantly higher than that of borderline ovarian tumor(27.8%) and benign ovarian epithelial tumor (18.9%), there was significantdifference (P<0.05); NFAT3and tissue differentiation, clinical stage and theabdominal organs and lymph node metastasis, differentiation is low, itsexpression is higher (P<0.05); Clinical staging of advanced rate higher thanthe early positive expression (P<0.05).The positive expression of abdominalorgans and lymph node metastasis were higher than those without metastasis(P<0.05), but not with ascites (P>0.05).(4)APR3, NFκB/p65, NFAT3expression in ovarian cancer tissue type and patient age were not related (P>0.05).2.APR3protein expression was positively correlated NFκB/p65protein(r=0.382, P <0.05)；but, between the APR3protein with NFAT3protein andbetween the NFκB/p65protein with NFAT3protein were notcorrelated (P<0.05)3.Effect of cisplatin on proliferation of human ovarian cancer SKOV3and3AO cells:in1μg/ml,5μg/ml,10μg/ml,20μg/ml and30μg/ml fiveconcentration range, cisplatin inhibition has a time and concentrationdependent on ovarian carcinoma SKOV3and3AO cell growth (P<0.05).4.Immunocytochemistry analysis APR3, NFκB/p65, NFAT3in ovariancancer SKOV3and3AO cells were expressed, mainly located in thecytoplasm. By cisplatin, the expression was decreased.5.Western blot analysis showed the cisplatin (10μg/ml) treatment ofovarian cancer SKOV3and3AO cells after24hours of the expression ofAPR3, NFκB/p65, NFAT3protein were weaked. before and after drug treatment the ovarian cancer SKOV3,3AO cells in APR3average relativeintensity of protein bands were0.976±0.042,0.406±0.012(P<0.05) and1.205±0.021,0.560±0.036(P<0.05); NFκB/p65average gray values ofprotein bands were1.094±0.074,0.954±0.046(P <0.05) and1.106±0.072,0.904±0.050(P <0.05); The average gray values of NFAT3proteinbands were0.614±0.046,0.135±0.004(P <0.05) and0.606±0.208,0.019±0.006(P <0.05);The relative gray value of APR3protein bands in humanovarian cancer cell of SKOV3compared with in3AO,the difference wasstatistically significant(P <0.05),while The relative gray value ofNFκ-B/p65protein bands and NFAT3protein bands in human ovarian cancercell of SKOV3compared with in3AO,the difference was no significant(P<0.05);In human ovarian cancer cell of SKOV3without cisplatin treatment,APR3expression between with NFκB/p65(r=0.949),APR3expressionbetween with NFAT3(r=0.985) and NFκB/p65expression between withNFAT3(r=0.988) were both positively correlated(P <0.05);In human ovariancancer cell of3AO without cisplatin treatment, APR3expression betweenwith NFAT3was positively correlated(r=0.981,P <0.05),but APR3expression between with NFκB/p65and NFκB/p65expression between withNFAT3were no correlation(P﹥0.05).Conclusion:1.Expression of APR3, NF κ B/p65, NFAT3protein in ovarian cancertissues were significantly higher than those in benign ovarian tumor.Expression of APR3, NF κ B/p65, NFAT3protein may be associated withovarian cancer. Significant correlation with APR3, NF κ B/p65, NFAT3protein and ovarian cancer tissue differentiation, clinical stage and theabdominal organs and lymph node metastasis, Over expression of APR3, NF κ B/p65, NFAT3protein, may be Involved in ovarian cancer, development andmetastasis.2.Cisplatin on ovarian cancer SKOV3,3AO cell proliferation weresignificantly inhibited, Cisplatin may through the expression of APR3, NF κB/p65and NFAT3protein, inhibit the proliferation of tumor cell.3.ExpressionAPR3protein expression and protein NFκB/p65are positively correlated inovarian cancer tissue,but in human ovarian cancer cell of SKOV3, APR3expression between with NFκB/p65,APR3expression between with NFAT3and NFκB/p65expression between with NFAT3were both positivelycorrelated(P <0.05);In human ovarian cancer cell of3AO, there are onlyAPR3expression between with NFAT3positively correlated,suggesting thatthe proteins of NF κ B/p65and NFAT3may enhance the expression of APR3promotes ovarian epithelial tumors.