Epithelial Ovarian Cancer Group Research Articles

Abstract LB381: Plasma microbiota as novel biomarkers of early epithelial ovarian cancer detection

Abstract Background: Ovarian cancer accounts for 2% of all female cancers in the United States yet it is the 5th leading cancer death among women. Advanced epithelial ovarian cancer (EOC) is associated with an overall survival rate of 30% but can be cured in up to 90% of cases if diagnosed early when the disease is still limited to the ovaries and fallopian tubes. While there is no effective screening method for EOC, developing novel and minimally invasive clinical tests is paramount to identify women with early disease as well as those who are at increased risk. Plasma-based microbiota signatures specific to EOC could become leading biomarkers of early disease in the field that complement existing clinical tools. Methods: We utilized an established institutional biobank to access 180 biobanked plasma samples from: (1) women with EOC; (2) women with non-EOC solid tumors; (3) women undergoing gynecologic-related procedures and non-EOC surgeries; and (4) healthy controls. Batch 1 included 96 plasma samples (n=24 per group) and Batch 2 included 84 samples (n=19 per group and 8 repeat EOC samples). Microbial profiles of plasma specimens were assessed using an amplicon-based sequencing method that targets the 16S rRNA gene in the bacterial genome to study bacterial phylogeny and taxonomy in diseased and healthy women. A stringent bioinformatic pipeline was applied to filter out suspicious bacterial contaminants at the genus level. Bacterial decontamination procedures in the pipeline included evaluating repeat plasma samples of EOC cases and sampling nucleic acid isolation kit reagents to serve as negative controls (only in Batch 2) . Differential abundances of the plasma microbiota among the groups were assessed using analysis of compositions of microbiomes (ANCOM) as implemented in R software package ANCOM-BC. Results: The 16S rRNA gene sequencing identified, classified, and quantified 564 bacterial genera in the circulation of women diagnosed with EOC, non-EOC solid tumors, benign gynecologic conditions, and healthy controls. Bioinformatic decontamination analysis removed bacteria: (1) with differing bacterial abundances between batches; (2) with higher bacteria in negative controls; (3) with differing bacterial abundances in repeat samples; and (4) known as contaminants in the literature that resulted in the removal of 225 high-risk bacterial contaminants. A total of 339 high quality bacteria remained for evaluation. Because we did not collect negative controls in Batch 1, only Batch 2 plasma samples (n=84; 27 in EOC group; 19 in each other group) were included in the final analysis. Women with EOC, which were primarily of the high-grade serous ovarian carcinoma histologic subtype had significantly distinct differential abundances of genera Brevibacterium (p <0.001), Chloronema (p <0.001), Facklamia (p <0.001), Zymomonas (p <0.001), and Sutterella (p <0.001) when compared to women with non-EOC solid tumors, benign gynecologic conditions, and healthy controls. Conclusions: These findings suggest that plasma EOC-associated bacteria could be exploited in the development of a microbial-based blood test for the diagnosis and early detection of EOC in routine practice settings. Citation Format: Diane E. Mahoney, Prabhakar Chalise, Dong Pei, Rachel Griffard, Trisha Home, Harsh Pathak, Shahid Umar, Andrew K. Godwin. Plasma microbiota as novel biomarkers of early epithelial ovarian cancer detection [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB381.

Fertility and Pregnancy Outcomes after Fertility-Sparing Surgery for Early-Stage Borderline Ovarian Tumors and Epithelial Ovarian Cancer: A Single-Center Study.

This study examined treatment outcomes, including preserved fertility, menstrual regularity, and pregnancy outcomes, in patients with stage I epithelial ovarian cancer (EOC) or borderline ovarian tumors (BOTs) who underwent fertility-sparing surgery (FSS). Patients with stage I EOC and BOTs who were aged 18-45 years and underwent FSS between 2007 and 2022 were retrospectively reviewed. Significant differences between various subgroups in terms of disease recurrence, menstrual irregularity due to the disease, and pregnancy outcomes were analyzed. A total of 71 patients with BOTs and 33 patients with EOC were included. In the BOT group, the median age was 30 (range, 19-44) years. Recurrence occurred in eight patients, with one case exhibiting a malignant transformation into mucinous EOC. Among the 35 married patients with BOTs, 20 successfully conceived, resulting in 23 live births and 3 spontaneous abortions. A higher pregnancy rate was observed in those without prior childbirth (82.4%) than in those who had prior childbirth (33.3%). In the EOC group, the median age was 34 (range, 22-42) years. Recurrence occurred in one patient. Menstrual regularity was maintained in 69.7% of the patients. Among the 14 married patients in this group, 12 achieved a total of 15 pregnancies (including 2 twin pregnancies), 16 live births, and 1 spontaneous abortion. The results of the study confirmed that FSS is a favorable surgical option for young women with early-stage BOTs or EOC who wish to preserve their fertility. However, additional investigations are needed to validate these findings.

Ascitic autotaxin as a potential prognostic, diagnostic, and therapeutic target for epithelial ovarian cancer

BackgroundMalignant ascites contributes to the metastatic process by facilitating the multifocal dissemination of ovarian tumour cells onto the peritoneal surface. However, the prognostic and diagnostic relevance of ascitic fluid remains largely unknown. Herein, we investigated the potential clinical value and therapeutic utility of ascitic autotaxin (ATX) in epithelial ovarian cancer (EOC).MethodsATX expression was assessed in clinical samples. Spheroid-forming assay, real-time PCR, western blot analysis, invadopodia assay, and adhesion assays were performed.ResultsAscitic ATX expression was highly elevated in patients with ovarian cancer compared to those with benign ascites and was associated with advanced stage, high grade, and a short disease-free period in patients with EOC. Combining the diagnostic ability of ascitic ATX and serum CA-125 levels significantly improved the area under the curve (AUC) value for EOC compared to serum CA125 level alone. This marker combination showed a large odds ratio for short disease-free period in high-risk EOC groups. Functional studies revealed that ascitic ATX was required for maintaining cancer stem cell-like characteristics and invadopodia formation.ConclusionAscitic ATX levels may serve as a useful prognostic indicator for predicting aggressive behaviour in EOC. ATX-linked invadopodia are a potential target to prevent peritoneal dissemination in ovarian cancer.

The performance of Carbohydrate Antigen 125-Thomsen-nouveau and anti-Müllerian hormone combined with CA125, Human epididymis protein 4 and Risk of Malignancy Algorithm in diagnosis for patients with Epithelial ovarian cancer

ObjectivesWe examined the blood concentrations of Carbohydrate Antigen 125-Thomsen-nouveau (CA125-Tn) and anti-Müllerian hormone (AMH) in epithelial ovarian cancer (EOC) patients to evaluate their potential diagnostic utility together with CA125, human epididymis protein 4 (HE4) and Risk of Malignancy Algorithm (ROMA). Design & methods50 healthy subjects, 45 EOC patients, 22 patients with borderline ovarian tumors (BOT), 21 patients with benign ovarian tumor (BET) and 45 patients with chocolate cyst of ovary (CCO) were studied. Blood levels of CA125, HE4, CA125-Tn and AMH were measured, and the ROMA value was calculated. We compared the differences in the levels of these biomarkers among groups. Additionally, a total of 10 testing strategies were established for comparison to maximize the diagnostic value. ResultsThe levels of CA125, HE4, CA125-Tn and ROMA value were significantly higher in EOC group compared with either the disease control (DC) group (BOT group, BET group and CCO group) or healthy control (HC) group (p < 0.001). In addition, they had better discriminatory performance with an area under the receiver operator characteristic curve (AUC) 0.93; 0.93; 0.93; 0.85, respectively (p < 0.001) compared with the AUC value of AMH 0.67 (p < 0.001). Among all 10 testing strategies, both single-positive of ROMA and double-positive of any 2 markers showed better Youden index (0.82, 0.79, respectively) and kappa value (κ) (0.82, 0.81, respectively). ConclusionsCA125-Tn and AMH can be treated as useful biomarkers of EOC when combined with CA125, HE4 and ROMA, because when any two biomarkers of them are positive, the value of EOC diagnosis is maximized.

Clinical Significance of the Combination of Serum HE4 Levels, Hemoglobin-to-Red Cell Distribution Width Ratio, and CT Imaging for the Pretreatment Assessment of Adnexal Masses.

Background: This study aimed to determine the optimal combination of biomarkers that can predict epithelial ovarian cancer (EOC) and compare the combination with the risk of ovarian malignancy algorithm (ROMA) or Copenhagen index (CPH-I). Methods: Data from 66 patients with EOC and 599 patients with benign ovarian masses who underwent definitive tissue diagnosis of adnexal masses between January 2017 and March 2021 were analyzed. The Mann-Whitney U test or Kruskal-Wallis test was used for between-group comparisons of medians. Logistic regression was used to establish an EOC predictor model. Area under the curve (AUC) comparisons between models were performed using the Delong nonparametric approach. Results: The median age of the patients was 43 years. Twenty-nine (43.9%) patients had early-stage disease (stages I-II) and 37 (56.1%) patients had advanced-stage disease (stages III-IV). The median age, body mass index, white blood cell count, hemoglobin-to-red cell distribution width ratio (HRR), platelet count, platelet-to-lymphocyte ratio, lymphocyte-to-monocyte ratio, serum albumin level, cancer antigen 125, human epididymal secretory protein 4 (HE4), ROMA, and CPH-I were significantly different between the stage I-IV EOC and benign ovarian mass groups. Multivariate logistic regression analysis revealed that HE4, HRR, and computed tomography (CT) imaging were significant predictors of both stages I-IV and I-II EOC. Using these covariates, an interim model (IM) (consisting of HE4 and HRR) and a full model (FM) (consisting of HE4, HRR, and CT imaging) were constructed. When predicting stage I-IV EOC, the AUC of IM was comparable to that of ROMA or CPH-I, whereas the AUC of FM outperformed ROMA or CPH-I. In predicting stage I-II EOC, the AUC of IM was comparable to that of CPH-I but higher than that of ROMA, and the AUC of FM outperformed ROMA or CPH-I. Conclusion: FM outperformed ROMA or CPH-I in predicting stage I-IV EOC and stage I-II EOC. Therefore, FM could be a promising model for improving preoperative prediction of EOC at an early stage. However, further prospective studies are required to validate these results.

Involvement of Cancer Stem Cells in Chemoresistant Relapse of Epithelial Ovarian Cancer Identified by Transcriptome Analysis.

Epithelial ovarian cancer (EOC) is the most lethal gynecological malignancy. Despite the initial resection and chemotherapeutic treatment, relapse is common, which leads to poor survival rates in patients. A primary cause of recurrence is the persistence of ovarian cancer stem cells (OCSCs) with high tumorigenicity and chemoresistance. To achieve a better therapeutic response in EOC relapse, the mechanisms underlying acquired chemoresistance associated with relapse-initiating OCSCs need to be studied. Transcriptomes of both chemosensitive primary and chemoresistant relapse EOC samples were obtained from ICGC OV-AU dataset for differential expression analysis. The upregulated genes were further studied using KEGG and GO analysis. Significantly increased expression of eighteen CSC-related genes was found in chemoresistant relapse EOC groups. Upregulation of the expression in four hub genes including WNT3A, SMAD3, KLF4, and PAX6 was verified in chemoresistant relapse samples via immunohistochemistry staining, which confirmed the existence and enrichment of OCSCs in chemoresistant relapse EOC. KEGG and GO enrichment analysis in microarray expression datasets of isolated OCSCs indicated that quiescent state, increased ability of drug efflux, and enhanced response to DNA damage may have caused the chemoresistance in relapse EOC patients. These findings demonstrated a correlation between OCSCs and acquired chemoresistance and illustrated potential underlying mechanisms of OCSC-initiated relapse in EOC patients. Meanwhile, the differentially expressed genes in OCSCs may serve as novel preventive or therapeutic targets against EOC recurrence in the future.

Develop a nomogram to predict overall survival of patients with borderline ovarian tumors.

BACKGROUNDThe prognosis of borderline ovarian tumors (BOTs) has been the concern of clinicians and patients. It is urgent to develop a model to predict the survival of patients with BOTs.AIMTo construct a nomogram to predict the likelihood of overall survival (OS) in patients with BOTs.METHODSA total of 192 patients with histologically verified BOTs and 374 patients with epithelial ovarian cancer (EOC) were retrospectively investigated for clinical characteristics and survival outcomes. A 1:1 propensity score matching (PSM) analysis was performed to eliminate selection bias. Survival was analyzed by using the log-rank test and the restricted mean survival time (RMST). Next, univariate and multivariate Cox regression analyses were used to identify meaningful independent prognostic factors. In addition, a nomogram model was developed to predict the 1-, 3-, and 5-year overall survival of patients with BOTs. The predictive performance of the model was assessed by using the concordance index (C-index), calibration curves, and decision curve analysis (DCA).RESULTSFor clinical data, there was no significant difference in body mass index, preoperative CA199 concentration, or tumor localization between the BOTs group and EOC group. Women with BOTs were significantly younger than those with EOC. There was a significant difference in menopausal status, parity, preoperative serum CA125 concentration, Federation International of gynecology and obstetrics (FIGO) stage, and whether patients accepted postoperative adjuvant therapy between the BOT and EOC group. After PSM, patients with BOTs had better overall survival than patients with EOC (P value = 0.0067); more importantly, the 5-year RMST of BOTs was longer than that of EOC (P value = 0.0002, 95%CI -1.137 to -0.263). Multivariate Cox regression analysis showed that diagnosed age and surgical type were independent risk factors for BOT patient OS (P value < 0.05). A nomogram was developed based on diagnosed age, preoperative serum CA125 and CA199 Levels, surgical type, FIGO stage, and tumor size. Moreover, the c-index (0.959, 95% confidence interval 0.8708–1.0472), calibration plot of 1-, 3-, and 5-year OS, and decision curve analysis indicated the accurate predictive ability of this model.CONCLUSIONPatients with BOTs had a better prognosis than patients with EOC. The nomogram we constructed might be helpful for clinicians in personalized treatment planning and patient counseling.

Circulating Exosomal miR-1290 for Diagnosis of Epithelial Ovarian Cancer.

The aim of the study was to develop a new diagnostic biomarker for identifying serum exosomal miRNAs specific to epithelial ovarian cancer (EOC) and to find out target gene of the miRNA for exploring the molecular mechanisms in EOC. A total of 84 cases of ovarian masses and sera were enrolled, comprising EOC (n = 71), benign ovarian neoplasms (n = 13). We detected expression of candidate miRNAs in the serum and tissue of both benign ovarian neoplasm group and EOC group using real-time polymerase chain reaction. Immunohistochemistry were constructed using formalin fixed paraffin embedded (FFPE) tissue to detect expression level of suppressor of cytokine signaling 4 (SOCS4). In the EOC group, miRNA-1290 was significantly overexpressed in serum exosomes and tissues as compared to benign ovarian neoplasm group (fold change ≥ 2, p < 0.05). We observed area under the receiver operating characteristic curve (AUC) for miR-1290, using a cut-off of 0.73, the exosomal miR-1290 from serum had AUC, sensitivity, and specificity values of 0.794, 69.2 and 87.3, respectively. In immunohistochemical study, expression of SOCS4 in EOC was lower than that in benign ovarian neoplasm. Serum exosomal miR-1290 could be considered as a biomarker for differential diagnosis of EOC from benign ovarian neoplasm and SOCS4 might be potential target gene of miR-1290 in EOC.

Clinical Significance of Tumor Infiltrating Lymphocytes in Association with Hormone Receptor Expression Patterns in Epithelial Ovarian Cancer.

Hormone receptor expression patterns often correlate with infiltration of specific lymphocytes in tumors. Specifically, the presence of specific tumor-infiltrating lymphocytes (TILs) with particular hormone receptor expression is reportedly associated with breast cancer, however, this has not been revealed in epithelial ovarian cancer (EOC). Therefore, we investigated the association between hormone receptor expression and TILs in EOC. Here we found that ERα, AR, and GR expression increased in EOC, while PR was significantly reduced and ERβ expression showed a reduced trend compared to normal epithelium. Cluster analysis indicated poor disease-free survival (DFS) in AR+/GR+/PR+ subgroup (triple dominant group); while the Cox proportional-hazards model highlighted the triple dominant group as an independent prognostic factor for DFS. In addition, significant upregulation of FoxP3+ TILs, PD-1, and PD-L1 was observed in the triple dominant group compared to other groups. NanoString analyses further suggested that tumor necrosis factor (TNF) and/or NF-κB signaling pathways were activated with significant upregulation of RELA, MAP3K5, TNFAIP3, BCL2L1, RIPK1, TRAF2, PARP1, and AKT1 in the triple dominant EOC group. The triple dominant subgroup correlates with poor prognosis in EOC. Moreover, the TNF and/or NF-κB signaling pathways may be responsible for hormone-mediated inhibition of the immune microenvironment.

The clinical value of the combined detection of sEGFR, CA125 and HE4 for epithelial ovarian cancer diagnosis.

This study aims to investigate the clinical value of combined detection of serum soluble epidermal growth factor receptor (sEGFR), cancer antigen 125 (CA125), and human epididymis protein 4 (HE4) in the diagnosis of epithelial ovarian cancer (EOC). From December 2017 to October 2018, the serum samples were obtained from the Affiliated Hospital of Xuzhou Medical University, with 30 patients as EOC group, 30 patients with benign ovarian neoplasms as benign group, and 17 healthy subjects as healthy group. Besides, among 30 EOC patients, 9 serum samples were obtained from pre-operative and post-operative EOC patients. The levels of serum sEGFR were detected by enzyme-linked immunosorbent assay (ELISA), while CA125 and HE4 were detected by enhanced chemiluminescence immunoassay (ECLIA). The diagnostic value was evaluated by receiver operating characteristic (ROC) curve analysis. The levels of serum sEGFR, CA125, and HE4 in EOC group were significantly higher than those in benign group (p<0.05) and healthy group (p<0.05). When using a single tumor marker, the CA125 shows the highest sensitivity (93.30%) and HE4 shows the highest specificity (97.87%). The specificity of combined detection of serum sEGFR, CA125, and HE4 was 100%, which was significantly higher than that using a single tumor marker. The area under the ROC curve (AUC) of combined detection of serum sEGFR, CA125, and HE4 (0.965) was much higher than that of the single detection and higher than that of combined detection of CA125 and HE4 (0.940). Moreover, the level of serum sEGFR in post-operative EOC group was significantly lower than that in the corresponding pre-operative EOC group (p<0.05). Our study shows that combined detection of serum sEGFR, CA125, and HE4 increases the specificity and efficiency in EOC diagnosis, indicating that sEGFR could be a potential biomarker for the diagnosis and prognosis of EOC.

HSPA1A, HSPA1L and TRAP1 heat shock genes may be associated with prognosis in ovarian epithelial cancer.

Epithelial ovarian cancer (EOC) is the most lethal gynecological malignancy, with the presence of chemoresistance contributing to the poor prognosis. Heat Shock Proteins (HSPs) genes are activated in response to pathophysiological stress and serve a role in a variety of stages in carcinogenesis, acting primarily as anti-apoptotic agents and in chemotherapy resistance in a variety of tumor types. The current study evaluated the HSP gene expression profile in women with ovarian cancer (OC) and their correlation with clinical and pathological aspects of patients with OC. A total of 51 patients included in the current study were divided into four groups: Primary Epithelial Ovarian Cancer (EOC; n=14), metastatic EOC (n=11), ovarian serous cystadenoma (n=7) and no evidence of ovarian malignancy or control groups (n=19). RNA extraction and reverse transcription-quantitative (RT-q) PCR was then performed on the samples obtained. RT-qPCR was performed to compare TNF receptor associated protein 1 (TRAP1), heat shock protein family (HSP) HSPB1, HSPD1, HSPA1A and HSPA1L expression in primary and metastatic EOCs. TRAP1, HSPB1, HSPD1, HSPA1A and HSPA1L gene expression did not differ among groups. HSPA1A, HSPA1L and TRAP1 were revealed to be underexpressed in the primary and metastatic EOC groups, with HSPA1L exhibiting the lowest expression. TRAP1 expression was higher in tumors at stages I/II compared with those at stages III/IV. No correlation was exhibited between HSP expression and age, menarche, menopause, parity, period after menopause initiation, cytoreduction, CA-125 or overall and disease-free survival. HSPA1A was negatively correlated with the risk of mortality from OC. The results indicated that the downregulation of HSPA1A, HSPA1L and TRAP1 could be associated with the clinical prognostic features of women with EOC.

Ovarian cancer circulating extracelluar vesicles promote coagulation and have a potential in diagnosis: an iTRAQ based proteomic analysis

BackgroundCirculating extracelluar vesicles (EVs) in epithelial ovarian cancer (EOC) patients emanate from multiple cells. These EVs are emerging as a new type of biomarker as they can be obtained by non-invasive approaches. The aim of this study was to investigate circulating EVs from EOC patients and healthy women to evaluate their biological function and potential as diagnostic biomarkers.MethodsA quantitative proteomic analysis (iTRAQ) was applied and performed on 10 EOC patients with advanced stage (stage III–IV) and 10 controls. Twenty EOC patients and 20 controls were applied for validation. The candidate proteins were further validated in another 40-paired cohort to investigate their biomarker potential. Coagulation cascades activation was accessed by determining Factor X activity.ResultsCompared with controls, 200 proteins were upregulated and 208 proteins were downregulated in the EOC group. The most significantly involved pathway is complement and coagulation cascades. ApoE multiplexed with EpCAM, plg, serpinC1 and C1q provide optimal diagnostic information for EOC with AUC = 0.913 (95% confidence interval (CI) =0.848–0.957, p < 0.0001). Level of activated Factor X was significantly higher in EOC group than control (5.35 ± 0.14 vs. 3.69 ± 0.29, p < 0.0001).ConclusionsOur study supports the concept of circulating EVs as a tool for non-invasive diagnosis of ovarian cancer. EVs also play pivotal roles in coagulation process, implying the inherent mechanism of generation of thrombus which often occurred in ovarian cancer patients at late stages.

Comprehensive Serum Glycopeptide Spectra Analysis (CSGSA): A Potential New Tool for Early Detection of Ovarian Cancer.

Objectives: To conduct a comprehensive glycopeptide spectra analysis of serum between cancer and non-cancer patients to identify early biomarkers of epithelial ovarian cancer (EOC). Methods: Approximately 30,000 glycopeptide peaks were detected from the digested serum glycoproteins of 39 EOC patients (23 early-stage, 16 advanced-stage) and 45 non-cancer patients (27 leiomyoma and ovarian cyst cases, 18 endometrioma cases) by liquid chromatography mass spectrometry (LC–MS). The differential glycopeptide peak spectra were analyzed to distinguish between cancer and non-cancer groups by employing multivariate analysis including principal component analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS-DA) and heat maps. Results: Examined spectral peaks were filtered down to 2281 serum quantitative glycopeptide signatures for differentiation between ovarian cancer and controls using multivariate analysis. The OPLS-DA model using cross-validation parameters R2 and Q2 and score plots of the serum samples significantly differentiated the EOC group from the non-cancer control group. In addition, women with early-stage clear cell carcinoma and endometriomas were clearly distinguished from each other by OPLS-DA as well as by PCA and heat maps. Conclusions: Our study demonstrates the potential of comprehensive serum glycoprotein analysis as a useful tool for ovarian cancer detection.

Association between AXIN1 gene polymorphisms and epithelial ovarian cancer in Chinese population.

Aim: The study was aimed to explore the association between AXIN1 gene polymorphism and epithelial ovarian cancer (EOC) susceptibility as well as prognosis. Methods: A total of 165 EOC cases and 327 healthy controls were recruited to participate in this study. In total three tag SNPs in AXIN1 gene were genotyped using PCR-restriction fragment length polymorphism. Results: Significantly increased EOC risk was found associated with the A/C heterozygous genotype of rs12921862, the C allele and C/T genotype in the rs1805105 and the T/T genotype of rs370681. Meanwhile, Kaplan-Meier survival curves showed that patients with rs12921862 C/C genotype improved overall survival in the EOC group. Conclusion: Our results suggest that the AXIN1 gene may be related to susceptibility and overall survival in EOC.

Potential of osteopontin in the management of epithelial ovarian cancer.

BackgroundOsteopontin (sOPN) is a promising blood tumour marker for detecting epithelial ovarian cancer (EOC). However, other clinical uses of sOPN as a tumour marker in EOC are still lacking. Since sOPN concentrations in serum are not associated with those in ascites, we compared clinical value of sOPN concentrations in the two body fluids.Patients and methodsThe study included 31 women with advanced EOC and 34 women with benign gynaecological pathology. In the EOC group, serum for sOPN analysis was obtained preoperatively, after primary debulking surgery and after chemotherapy. In the control group, serum was obtained before and after surgery. Ascites and peritoneal fluid were obtained during surgery. sOPN concentrations were determined by flow cytometry bead-based assay.ResultsThe sensitivity and specificity of sOPN in detecting EOC was 91.2% and 90.3% (cut-off = 47.4 ng/ml) in serum, and 96.8% and 100% (cut-off = 529.5 ng/ml) in ascites. Kaplan-Meier analysis showed a significant association between higher serum sOPN concentration and overall survival (p = 0.018) or progression free survival (p = 0.008). Higher ascites sOPN concentrations were associated with suboptimally debulked tumour and unresectable disease. Higher serum sOPN concentrations were associated with refractory disease or incomplete response to platinum-based chemotherapy.ConclusionsThe study showed that ascites sOPN level mirrors present disease and is superior to serum level for diagnostic purposes and surgical planning, although the end result of treatment is the response of the whole body in fighting the disease. The preoperative sOPN concentration in serum thus better reflects disease outcome.

TMEFF1 overexpression and its mechanism for tumor promotion in ovarian cancer.

BackgroundTransmembrane protein with epidermal growth factor-like and two follistatin-like domains 1 (TMEFF1) has an anticarcinogenic effect in brain tumors. However, little is known about the role of TMEFF1 in epithelial ovarian cancer (EOC).Materials and methodsTMEFF1 expression in EOC was detected by immunohistochemistry; its relationship with clinical pathological parameters and its influence on prognosis were analyzed. The MTT, scratch, Transwell assays, and flow cytometry were used to assess the malignant behavior of ovarian cancer cell. Changes in node proteins in MAPK and PI3K/AKT signaling pathways and the expression of epithelial–mesenchymal transformation markers were measured by Western blot. The regulatory effect of p53 on TMEFF1 was verified by chromatin immunoprecipitation (ChIP) assay and Western blot.ResultsTMEFF1 expression was higher in the EOC group than in the borderline and benign tumor groups and normal ovary group; its high expression was significantly related to International Federation of Gynecology and Obstetrics stage (P=0.024) and independently predicted shorter overall survival (P<0.01). TMEFF1 overexpression in ovarian cancer cells induced increased cellular proliferation, migration, and invasion but reduced apoptosis. In addition, the percentage of phosphorylated node proteins in MAPK and PI3K/AKT signaling pathways increased significantly. The expression of E-cadherin decreased but that of vimentin and N-cadherin increased. After the addition of MAPK (PD98059) and PI3K (GDC-0941) pathway inhibitors, ovarian cancer cells overexpressing TMEFF1 showed suppressed malignant behavior. TMEFF1 protein expression in an ovarian cancer cell lines (CAOV3 and ES-2) was downregulated after the inhibition of TP53. The transcription factor, p53, bound the promoter region of the TMEFF1 gene according to ChIP.ConclusionTMEFF1 is a carcinogenic gene in ovarian cancer and can be regulated by p53 transcription. Through MAPK and PI3K/AKT signaling pathways, TMEFF1 promotes the malignant behavior in EOC. Therefore, TMEFF1 may be considered as a potential therapeutic target for ovarian cancer.

Comparison of stage III mucinous and serous ovarian cancer: a case-control study

BackgroundThe purpose of this case-control study was to compare the prognoses of women with stage III mucinous ovarian carcinoma (MOC) who received maximal or optimal cytoreduction followed by paclitaxel plus carboplatin chemotherapy to those of women with stage III serous epithelial ovarian cancer (EOC) treated in the similar manner.MethodsWe performed a multicenter, retrospective review to identify patients with stage III MOC at seven gynecologic oncology departments in Turkey. Eighty-one women with MOC were included. Each case was matched to two women with stage III serous EOC in terms of age, tumor grade, substage of disease, and extent of residual disease. Survival estimates were measured using Kaplan-Meier plots. Variables predictive of outcome were analyzed using Cox regression models.ResultsWith a median follow-up of 54 months, the median progression-free survival (PFS) for women with stage III MOC was 18.0 months (95% CI; 13.8–22.1, SE: 2.13) compared to 29.0 months (95% CI; 24.04–33.95, SE: 2.52) in the serous group (p = 0.19). The 5-year overall survival rate of the MOC group was significantly lower than that of the serous EOC group (44.9% vs. 66.3%, respectively; p < 0.001). For the entire cohort, presence of multiple peritoneal implants (Hazard ratio [HR] 2.39; 95% confidence interval [CI], 1.38–4.14, p = 0.002) and mucinous histology (HR 2.28; 95% CI, 1.53–3.40, p < 0.001) were identified as independent predictors of decreased OS.ConclusionPatients with MOC seem to be 2.3 times more likely to die of their tumors when compared to women with serous EOC.

Expression of human epididymal protein 4 in patients with epithelial ovarian cancer and its effect on prognosis

Objective To investigate the expression of human epididymal protein 4 (HE4) and its effect on prognosis in patients with epithelial ovarian cancer (EOC). Methods From 2009 to 2014, ninety-eight cases of epithelial ovarian cancer (EOC group) and 29 cases of benign ovarian tumor (benign group) and 100 healthy subjects (control group) were selected as research subjects.The serum HE4 was detected by double antibody sandwich enzyme-linked immunosorbent assay (ELISA), and the positive expression was determined by HE4>150 pmol/L in serum.The concentration and positive rate of serum HE4 were compared among the three groups.The median serum HE4 level of EOC patients before treatment was divided into high level HE4 group and low level HE4 group.The median survival time of the two groups of EOC patients was compared. Results The median value of serum HE4 concentration (four percentile) in EOC group was 421.8 (68.7 ~ 2173.9) pmol/L, 47.8 (35.24 ~ 68.5) pmol/L in the benign group and 43.8 (32.91 ~ 70.4) pmol/ L in the control group.The serum concentration of HE4 in the EOC group was significantly higher than that in the benign group and the control group (P=0.000). The serum HE4 concentration in group EOC was significantly higher than that in benign group and control group (P 0.05). The positive rate of serum HE4 in the EOC group was 74.5 (73/98), the positive rate of the benign group and the control group were 0, the difference between the three groups was statistically significant (P=0.000). The positive rate of serum HE4 in the EOC group was significantly higher than that in the benign group and the control group (P 0.05). The median survival time of EOC patients in low level HE4 group was 66.2 months, while that in high level HE4 group was 36.9.The difference between two groups was statistically significant (P=0.001). Conclusion The serum HE4 level of EOC patients was significantly higher than that of patients with benign ovarian tumors and healthy controls.The higher serum HE4 level, the worse the prognosis, and the shorter the survival time of patients.Therefore, the expression level of serum HE4 can be used for the diagnosis and prognosis of ovarian cancer. Key words: Epithelial ovarian cancer; human epididymis protein 4; carbohydrate antigen 125; survival time

Characteristics of the Distribution of Ferritin in Epithelial Ovarian Tumor Patients: Results of a Retrospective, Observational Study

Purpose: To investigate the ferritin distribution in epithelial ovarian cancer patients according to the FIGO stage in the prognosis of epithelial ovarian cancer. Method: All ovarian cancer patients were divided into two groups according their FIGO stage. Benign ovarian tumor patients were analyzed as the control. Serum ferritin, serum iron, and other related medical index were detected by automatic instruments for all patients. In addition, ferritin heavy chain (FHC) and ferritin light chain (FLC) proteins were detected by immunohistochemical staining in 60 epithelial ovarian cancer (EOC) patients and 30 benign ovarian tumor (BOT) patients, which were diagnosed in our department between 2011 and 2016. Results: The serum ferritin concentration was significantly higher in the EOC group than in the BOT group (172.56 ± 99.39 ng/mL vs 78.18 ± 43.06 ng/mL; p μmol/L vs 14.92 ± 6.36 μmol/L; p p p p p p p > 0.05). Conclusion: Patients showed an overexpression of ferritin and a downregulation of serum iron correlated with the prognosis of epithelial ovarian cancer, which may be a new target for diagnosis and treatment of epithelial ovarian cancer.

Suppression of endothelial cell migration by tumor associated macrophage-derived exosomes is reversed by epithelial ovarian cancer exosomal lncRNA

ObjectiveTo study the mechanism by which epithelial ovarian cancer (EOC)-derived exosomes restore the migration of endothelial cells that is suppressed by TAM-derived exosomes.MethodsExosomes were isolated from TAMs in the ascites of patients with EOC. The effect of exosomes on the expression of endothelial cell miRNA was monitored by PCR. The miRNA mimics were transfected to explore their effects. Microarray data and literature searches were used to predict target genes and the impact of target gene pathways, and small interfering RNA was used to target these genes. We used migration assays to determine whether ovarian cancer cell-derived exosomes participate in the regulation of TAMs and endothelial cells. We used microarray data to identify the target lncRNA, and we constructed target lncRNA expression plasmids to validate targets by Western blotting.ResultsWe separated TAMs from the ascites of patients with EOC and isolated exosomes from TAM supernatants. After co-culture with HUVECs, these exosomes were efficiently incorporated into HUVECs. The migration of HUVECs was suppressed significantly in the exosome group compared with blank controls (P < 0.05).The miRNA mimic transfection and target gene prediction found that TAM-derived exosomes targeted the miR-146b-5p/TRAF6/NF-κB/MMP2 pathway to suppress endothelial cell migration; this result was supported by PCR and Western blotting analyses. The expression of exosomal miR-146b-5p isolated from serum in the EOC group was significantly increased compared to healthy individuals. Finally, TAM-derived exosomes and EOC SKOV3-derived exosomes in combination stimulated HUVEC cells and overcame the inhibition of endothelial cell migration caused by TAM-derived exosomes. Two lncRNAs that were carried by SKOV3-derived exosomes were identified as NF-κB pathway-associated genes by Western blotting.ConclusionTAM-derived exosomes can inhibit the migration of endothelial cells by targeting the miR-146b-5p/TRAF6/NF-kB/MMP2 pathway. However, EOC-derived exosomes can transfer lncRNAs to remotely reverse this effect of TAMs on endothelial cells.