| ABSTRACTObjective: Female reproductive system includes three types: ovarian carcinoma, cervical cancer, and endometrial cancer. According to a survey in 2013, the American Cancer Society, in the three malignant tumors, ovarian carcinoma is a leading cause of death as a result of the deficiency of the molecular pathogenesis. Ovarian epithelial cancer includes a varied of histology types and there are marked differences among those types. We elected high-grade ovarian serous adencarcinoma which was the most common type in ovarian serous adencarcinoma as study object, it could eliminate the influences of histology type and grade on prognosis, more objectively reflect oncogenesis and progression of carcinoma, and provide evidences to exactly judge the clinical prognosis.According a new study, a hallmark of ovarian serous carcinoma is defective homologous DNA repair, at the same time APE1/Ref-1 and NPM1 play two important roles in DNA repair mechanisms. Recent studies have discovered the molecular interaction between NPM1 and APE1/Ref-1, which modulates a new function of APE1/Ref-1 in r-RNA metabolism. This interaction involves Lys27/31/32/35 residues of APE1/Ref-1 and the oligomerization domain of NPM1. Observations prove that NPM1 has a significant stimulatory effect on APE1/Ref-1 DNA repair activity in BER and that without this interaction, the proliferation of tumor cell lines is affected.In this study, high-grade ovarian serous andenocarcinoma tissues were selected as experimental group; normal fallopian tube tissues were selected as control group. The variance in APE1/Ref-1 and NPM1 expression were detected with immunohistochemistry staining. On the base, we explore the relationship in APE1/Ref-1, NPM1 expression and pathogenesis, progression, prognosis factors, and access prediction value of each prognosis factor, judge the independent prognosis factors.Methods:1 Research object:We collect 50 cases of high-grade ovarian serous carcinoma with complete clinical information, follow-up data, surgical resection and exact pathology from the Second Hospital of Hebei Medical University form July 2003 to January 2009. 20 normal fallopian tube tissues which were cutted because of other disease were selected as control.2 ImmunohistochemistryWe use three-steps method of immunohistochemistry to detect the expression of APE1/Ref-1 、NPM1 and PCNA in high-grade ovarian serous carcinoma and normal fallopian tube tissues. Negative control was carried out in identical condition excluding PBS insteading of antibody. Positive controls use the tissue which has been known that the target gene has the strong expression.3 Survival analysis50 high-grade serous ovarian carcinoma patients with follow-up were analyzed and made a summary. 36 and 60 months were cut-off point. According to compare the sum survival rates of 3 years and 5 years survival rates to find prognostic factors and calculate the risk coefficient. So we can find the independent factors of high-grade serous ovarian adenocarcinoma.4 Statistical treatmentsIn this study we use SPSS13.0 statistical package to analysis the statistics. X2 text was used in enumeration data. Mann-Whitney U text was used in ranked data. Spearman rank correlation was used to analysis the correlation of all the indexes. Kaplan-Meier univariate survival analysis was used to calculating the survival rate. Log-Rank test was used to comparing the different of survival rate. Cox multivariate survival analysis was used to compare the effect on prognosis of degree, judge the independent predictive factors. α=0.05.Rusults: 1 The expression of APE1/Ref-1、NPM1 and PCNA in high-grade ovarian serous adenocarcinoma 1.1 APE1/Ref-1 protein expressionThe result of IHC showed that the positive rates of APE1/Ref-1 protein expression in high-grade ovarian serous adenocarcinoma were 98%(cytoplasm expression as positive expression), significantly higher than that 0% in normal fallopian tube tissues(P<0.05). 1.2 NPM1 protein expressionThe result of IHC showed that the positive rates of NPM1 protein expression in high-grade ovarian serous adenocarcinoma were 96%(cytoplasm expression as positive expression), significantly higher than that 5% in normal fallopian tube tissues(P<0.05). 1.3 PCNA protein expressionThe result of IHC showed that the high expression rates(PCNA index >40%) of NPM1 protein expression in high-grade ovarian serous adenocarcinoma were 68%(nucleus expression as positive expression), significantly higher than that 10% in normal fallopian tube tissues(P<0.05). 2 The relationship between APE1/Ref-1, NPM1 and clinicpathological characteristics in high-grade ovarian serous adenocarcinomaThe significantly differences of APE1/Ref-1, NPM1 protein expression were not found in age, family history and ascites(P>0.05). The significantly differences of APE1/Ref-1, NPM1 protein expression were found in lymphatic metastasis, chemoresistance, FIGO stage, and distant metastasis(P<0.05), and the expression level in those tissue with lymphatic metastasis, chemoresistance, advanced stage, distant metastasis was higher than those tissue with not lymphatic metastasis, chemoresistance, distant metastasis and early stage. 3 The correlation between APE1/Ref-1, NPM1 and PCNA protein in high-grade ovarian serous adenocarcinoma 3.1 The correlation between APE1/Ref-1 and PCNA protein expression32 cases were high expression with APE1/Ref-1 in 50 high-grade ovarian serous adenocarcinoma, among, 28 cases were high expression with PCNA. Statistically, positive correlation was found between APE1/Ref-1 and PCNA protein expression in high-grade ovarian serous adenocarcinoma(r=0.797 P<0.05). 3.2 The correlation between NPM1 and PCNA protein expression33 cases were high expression with NPM1 in 50 high-grade ovarian serous adenocarcinoma, among, 31 cases were high expression with PCNA. Statistically, positive correlation was found between NPM1 and PCNA protein expression in high-grade ovarian serous adenocarcinoma(r=0.736 P<0.05). 3.3 The correlation between NPM1 and APE1/Ref-1 protein expression32 cases were high expression with APE1/Ref-1 in 50 high-grade ovarian serous adenocarcinoma, among, 28 cases were high expression with NPM1 Statistically, and positive correlation was found between NPM1 and APE1/Ref-1 protein expression in high-grade ovarian serous adenocarcinoma(r=0.730 P<0.05). 4 The effect of APE1/Ref-1 and NPM1 protein expression on prognosis in high-grade ovarian serous adenocarcinoma.According to the result of IHC and follow-up date, the survival rates of 3 and 5 years were 18.79% and 0% in high-grade ovarian serous adenocarcinoma with APE1/Ref-1 high expression, significantly lower than 83.33% and 38.89% in patients with APE1/Ref-1 low expression.According to the result of IHC and follow-up date, the survival rates of 3 and 5 years were 15.63% and 0% in high-grade ovarian serous adenocarcinoma with PCNA high expression, significantly lower than 88.89% and 38.89% in patients with PCNA low expression. 5 COX multivariate survival analysis of high-grade ovarian serous adenocarcinomaKaplan-Meier univariate survival analysis and Log-Rank test showed lymphatic metastasis, chemoresistance, FIGO stage, distant metastasis, APE1/Ref-1 expression and NPM1 expression were prognosis factors of patients with high-grade ovarian serous adenocarcinoma.COX multivariate survival analysis showed NPM1 expression was a independent prognosis factor, followed by APE1/Ref-1 expression.Conclusion:1 According to the result of IHC, altered subcellular APE1/Ref-1 and NPM1 expression were found in high-grade ovarian serous adenocarcinoma. It suggested that APE1/Ref-1 and NPM1 perform a significant role in oncogenesis of high-grade ovarian serous adenocarcinoma.2 The significantly differences of APE1/Ref-1, NPM1 protein expression were found in lymphatic metastasis, chemoresistance, FIGO stage, and distant metastasis. It suggested that APE1/Ref-1 and NPM1 may perform a significant role in progression, aggressive, and chemoresistance of high-grade ovarian serous adenocarcinoma.3 Positive correlations were found between APE1/Ref-1, NPM1 and PCNA protein expression in high-grade ovarian serous adenocarcinoma. It suggested that APE1/Ref-1 and NPM1 may perform a significant role in cell proliferation.4 Positive correlations were found between NPM1 and APE1/Ref-1 protein expression in high-grade ovarian serous adenocarcinoma. It suggested that APE1/Ref-1 interacts with NPM1 to contribute to oncogenesis and progression of high-grade ovarian serous adenocarcinoma.5 Beside conventional prognosis factors, APE1/Ref-1 and NPM1 were prognosis factors, and NPM1 expression was an independent prognosis factor, followed by APE1/Ref-1 expression. The two genes will be the new target of gene therapy. |
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