Study On Biomarkers For Low-and High-grade Serous Ovarian Carcinomas Clinical Analysis Of Ovarian Preservation For Endometrial Carcinomas In Women Aged40Years And Younger

Part Ⅰ:Study on Biomarkers for Low-and High-grade Serous Ovarian Carcinomasbackground:Ovarian serous carcinoma (OSC) is the most common histologic type of epithelial ovarian cancer. In2004, Malpica et al described a novel two-tier grading system for grading OSC as either low-grade serous carcinoma (LGSC) or high-grade serous carcinoma (HGSC). It has been found that the differences between the low-and high-grade cases are not limited to the pathology, but also detected at the origin, molecular pathogenic, as well as in the clinical features, especially in the aspect of chemo-sensitivity and prognosis. Therefore, the two-tier grading system has allowed us to further understand the development mechanism of OSC. It is imminent to search for specific biomarkers for LGSC and HGSC, and provide theoretical basis for molecular targeted therapy for OSC patients. Objective:To analyze the clinicopathological characteristics and prognosis of LGSC and HGSC. To search for specific molecules expressed in LGSC and HGSC and validate candidate makers in clinical samples, and to find new targets for individual treatment of OSC.Methods:The pathologist reviewed the pathology of6cases of primarily cultured ovarian cancer and selected patients with OSC, and classified those cases according to the two-tier grading system of MDACC. And then, the ovarian cancer-derived secretory/releasing proteome database was analyzed with DAVID and R Software. After that, candidate proteins were selected and their levels in the plasma samples from265OSC patients and102healthy controls were measured by double antibody sandwich enzyme-linked immunosorbent assay. The candidate proteins levels in the ovarian tumor tissues were measured by immunohistochemistry (IHC). In addition, this study validates the preliminary observations and explores the clinical significance of NID1in a large sample size of OSC patients.Results:(1). Clinical and pathological recording of271cases of OSC patients were retrospectively analyzed, they were divided into two groups:LGSC (n=38,12.7%) and HGSC (n=233,87.3%). There were no significant differences in the family history, preoperative level of CA125, surgical staging, lymphonode metastasis, neoadjuvant chemotherapy, lymphadenectomy and postoperative treatment between two groups (all P>0.05). The proportion of patients with age younger than50years in group LGSC was less than in HGSC group, the difference was statistically significant (P=0.027), while there was no statistical significant in the proportion of patients who was platinum-resistant between two groups (P=0.280). Compared with the WHO system,57.1%of grade1and grade2fall into the LGSC category, while98.6%of grade2and grade3fall into the high-grade category. The median follow-up time was26.9months (range:2.4to79.9months). Kaplan-Meier analysis revealed that two-tier grading system has significant effect on OS (P=0.034), but has no effect on PFS (P=0.225). Multivariate analysis showed that the two-tier grading system was not an independent prognostic factor for OSC.(2). Two candidate proteins (β-catenin and Grb2) were selected from the database for validation. Their levels in the plasma of OSC patients and healthy controls were measured by ELISA. We found that the plasma level of β-catenin and Grb2in OSC patients was significantly lower than that of healthy cohort (P<0.001). There were not, however, noticeable correlations between plasma levels of β-catenin or Grb2with age of onset, residual tumor size, FIGO stage, grade (WHO system), two-tier grading system, number of lymphonode metastasis, chemosensitivity and prognosis of OSC patients investigated (all P>0.05). In the group of patients received neoadjuvant chemotherapy, the level of Grb2was significantly higher than the patients who underwent primary debulking surgery (P<0.05).(3). The protein levels p53, β-catenin and Grb2in the ovarian tumor tissues were measured by IHC, especially in OSC patients. There was a statistically significant expression of p53and Grb2in LGSC compared with HGSC (P<0.05). In LGSC, the positive rate of p53is lower, with scattered strong positive cells, and higher positive rate of Grb2; however, in HGSC, the positive expression of p53was higher, with diffuse strong positive expression, and lower positive rate of Grb2;the protein level of β-catenin in OSC tissues has no correlation with the two-tier grading system, and the patients with positive expression of P-catenin protein had a better prognosis than those with negative expression. There were no correlations between levels of p53, β-catenin or Grb2in the OSC tissues with age of onset, family history, NAC, the residual tumor size, FIGO stage, or grade (WHO system)(all P>0.05).(4). We found that protein levels of NID1were considerably raised in the plasma from OSC patients compared to those in healthy controls, especially elevated in patients with advanced-stage (stage Ⅲ/Ⅳ) and those received neoadjuvant chemotherapy (P<0.05), However, it was irrelevant to residual tumor size, the grade (WHO system), two-tier grading system and chemotherapy sensitivity of the OSC cases investigated (all P>0.05). ROC curve analysis for NID1showed that it could discriminate patients with OSCs from healthy controls (AUC:0.65,95%CI,0.59-0.71), but not for the early diagnosis of (AUC0.65,95%CI,0.41～0.54), and CA125was superior.Conclusions:(1). Compared with HGSC, the incidence of LGSC is low, age of onset is younger, and the prognosis is well. Two-tier grading system was not an independent prognostic factor for OSC.(2). It is unsuitable for β-catenin or Grb2to be serum biomarker for ovarian serous cancer.(3). There was a statistically significant expression of p53and Grb2in LGSC compared with HGSC. Patients with positive expression of β-catenin protein had a better outcome than those with negative expression.(4). Plasma NID1may be used as a diagnostic biomarker for OSC and reflect the tumor burden. Part II:Clinical Analysis of Ovarian Preservation for Endometrial Carcinomas in Women Aged40Years and YoungerBackground:In recent years, along with the increasing incidences of endometrial carcinoma, the prevalence rate in young patients assumes the trend of escalation. The main surgical procedures of endometrial carcinoma are hysterectomy and bilateral salpingo-oophorectomy. However, it will seriously affect the quality of life of young patients with their ovaries removed, and increase the overall mortality rate. Therefore, in order not to increase the risk of death for young women with early stage endometrial carcinoma, how to preserve the function of ovary was becoming a hot topic.Objective:To investigate the clinical characteristics and prognosis of endometrial carcinomas in women aged40years or younger, and analyzes the safety of ovarian preservation for stage I patients.Methods:140cases of endometrial cancer aged younger than40from Jan1999to Jan2012were treated in Cancer Hospital, Chinese Academy of Medical Sciences. Seventy-five cases of Stage I endometrial cancer were further divided into the following two groups:20patients who underwent ovarian preservation (group A) and55patients who underwent oophorectomy (group B). Clinical and pathological recordings of these patients were reviewed and the two groups were compared.Results:(1). The study group constituted7.9%of all the endometrial cancer inpatients admitted to our hospital in the same period. FIGO Stage Ⅰ-Ⅱ accounted for73.6%, high and moderate differentiation cases did for82.9%, and88.6%of the cases were endometrioid adenocarcinoma. The5-years overall survival and disease-free survival rates for the whole group were90.3%,84.8%, respectively. Multivariate regression analysis displayed that ovarian involvement was the independent prognostic factors for the overall survival and disease-free survival rates (P<0.05). Coexisting ovarian malignancy was detected in26(18.6%) of140patients.Factors predictive of coexisting ovarian malignancy by multivariate analysis were ranked in the following order according to risk intensity:parametrial invasion, the level of CA125, surgical staging and age.(2). In the group A, there were13patients preserved both ovaries, and7patients preserved a single ovary. There were no significant differences in the age, body mass index, surgical staging, histology, grade, and cytology of peritoneal lavage or ascites, postoperative treatment between these two groups (all P>0.05). The differences in the level of CA125[25%(5/20) versus18%(10/55)] and number of patients underwent pelvic lymphadenectomy [35%(7/20) versus84%(46/55)] were statistically significant between the two groups (all P<0.05). Of seventy-five cases, only two patients relapsed and all survived after a median follow-up of31.7months (range from0to160months). Kaplan-Meier analysis revealed no difference in overall survival (100%versus100%) and disease free survival (90.0%versus95.5%) between two groups (P=0.579).Conclusions:Endometrial cancer of younger women is mostly lower-stage, endometrioid and has a better prognosis, while prognosis is poor when coexisting with ovarian tumors. Ovarian preservation has no statistically significant impact on the survival of young patients with stage I, well differentiated endometrial cancer. Large-scale, prospective clinical studies are needed to validate the safety of ovarian preservation for those patients.