Ovarian Serous Adenocarcinoma Research Articles

The diagnostic value of the p16ink marker for verification of tumors of unknown primary site in women with isolated lesion of inguinal lymph nodes.

Carcinomas with an unknown primary site are a heterogeneous group of metastatic tumors, numbering from 3% to 5% of all malignant carcinoma phenotypes. An isolated lesion of the inguinal lymph nodes is relevantly associated with primary localization of tumors in the pelvis, anal canal, lower abdomen, lower limbs, and more towards in the reproductive system (vulva, vagina and cervix for women, penis for men). But in the absence of clinical signs of cancer after careful research, the histological biopsy of the lymph node with additional immunohistochemical staining with organ-specific markers becomes the diagnostic method of choice. A retrospective analysis of the histological, morphometric and immunohistological characteristics of the biopsy material of 59 patients with isolated lesions of the inguinal lymph nodes (35 women and 24 men) aged from 20 to 87 years (mean 59.26±15.86; median 62 years) was conducted. The distribution of variants of an isolated lesion of the inguinal lymph nodes in women showed 14 of 35 (40.00%) metastatic tumors (the other 21 are lymphoproliferative conditions that require phenotyping, but not finding the primary localization). Of 14 metastatic lesions in women, 6 cases demonstrated a p16 ink (+) phenotype. For comparison, of 24 cases in men, only 6 (25.00%) were of metastatic origin, of which p16 ink (+/-) was partially 1 metastasis of melanoma (16.67%). After a thorough immunohistochemical study with a panel of organo-specific markers among isolated metastatic lesions of the inguinal lymph nodes of tumors in women, half of the revealed localized sites (7 out of 14) were in reproductive organs, namely: 4 p16 ink (+) to squamous cervical cancers, 2 p16 ink (+) to serous ovarian adenocarcinoma, 1 p16 ink (-) to leiomyosarcoma of the uterus. Other localizations did not have a clear location, as they related to metastases of carcinoma from Merkel cells and melanomas, as a result of frequent reduction of the primary tumor lesion in the skin.

Cancer stem cells contribute to angiogenesis and lymphangiogenesis in serous adenocarcinoma of the ovary.

The origin of blood and lymphatic vessels in high-grade serous adenocarcinoma of ovary (HGSOC) is uncertain. We evaluated the potential of cancer stem cells (CSCs) in HGSOC to contribute to their formation. Using spheroids as an in vitro model for CSCs, we have evaluated their role in primary malignant cells (PMCs) in ascites from previously untreated patients with HGSOC and cell lines. Spheroids from PMCs grown under specific conditions showed significantly higher expression of endothelial, pericyte and lymphatic endothelial markers. These endothelial and lymphatic cells formed tube-like structures, showed uptake of Dil-ac-LDL and expressed endothelial nitric oxide synthase confirming their endothelial phenotype. Electron microscopy demonstrated classical Weibel-Palade bodies in differentiated cells. Genetically, CSCs and the differentiated cells had a similar identity. Lineage tracking using green fluorescent protein transfected cancer cells in nude mice confirmed that spheroids grown in stem cell conditions can give rise to all three cells. Bevacizumab, a monoclonal antibody that targets vascular endothelial growth factor inhibited the differentiation of spheroids to endothelial cells in vitro. These results suggest that CSCs contribute to angiogenesis and lymphangiogenesis in serous adenocarcinoma of the ovary, which can be inhibited.

Robustness of RNA sequencing on older formalin-fixed paraffin-embedded tissue from high-grade ovarian serous adenocarcinomas.

Formalin-fixed paraffin-embedded (FFPE) tissues are among the most widely available clinical specimens. Their potential utility as a source of RNA for transcriptome studies would greatly enhance population-based cancer studies. Although preliminary studies suggest FFPE tissue may be used for RNA sequencing, the effect of storage time on these specimens needs to be determined. We conducted this study to determine whether RNA in archived FFPE high-grade ovarian serous adenocarcinomas from Surveillance, Epidemiology and End Results (SEER) registries was present in sufficient quantity and quality for RNA-Seq analysis. FFPE tissues, stored from 7 to 32 years, were obtained from three SEER sites. RNA was extracted, quantified, quality assessed, and subjected to RNA-Seq (a whole transcriptome sequencing technology). FFPE specimens stored for longer periods of time had poorer RNA sample quality as indicated by negative correlations between specimen storage time and fragment distribution values (DV). In addition, sample contamination was a common issue among the RNA, with 41 of 67 samples having 5% to 48% bacterial contamination. However, regardless of specimen storage time and bacterial contamination, 60% of the samples yielded data that enabled gene expression quantification, identifying more than 10,000 genes, with the correlations among most biological replicates above 0.7. This study demonstrates that FFPE high-grade ovarian serous adenocarcinomas specimens stored in repositories for up to 32 years and under varying storage conditions are a promising source of RNA for RNA-Seq. We also describe certain caveats to be considered when designing RNA-Seq studies using archived FFPE tissues.

The expression and clinical significance of YAP and cIAP1 in ovarian serous adenocarcinoma

The expression and clinical significance of YAP and cIAP1 in ovarian serous adenocarcinoma

Highlights from Recent Cancer Literature

Highlights from Recent Cancer Literature

Suppression of autophagy enhances preferential toxicity of epothilone A and epothilone B in ovarian cancer cells

BackgroundEpothilones are microtubule-targeting agents that induce death in a variety of cancer cell types. Here, we focus on the cellular and molecular mechanisms underlying epothilone A (Epo A) and epothilone B (Epo B)-induced autophagy and apoptosis in ovarian cancer cells, compared to the actions of the widely used clinical chemotherapy drug paclitaxel (PTX). Materials and methodsAutophagy was examined in two cell lines, SKOV-3 (human ovarian adenocarcinoma) and OV-90 (human ovarian papillary serous adenocarcinoma), which differ in the levels of p-glycoprotein and drug resistance, based on the LC3 ELISA assay, fluorescence detection of autophagosome formation, morphological changes evaluated via acridine orange staining, and visualization of LC3 protein using confocal microscopy. Cell viability was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay. Apoptosis was measured via the caspase-3/7 assay and immunofluorescence labeling of caspase-3. Differences in microtubule organization in epothilone-treated cells were investigated using specific antibodies against β-tubulin. All probes were analyzed both in the presence and absence of the autophagy inhibitor, bafilomycin A1 (Baf), and apoptosis inhibitor, Z-FA-FMK. ResultsEpothilone and PTX treatment induced a dose-dependent decrease in cell viability, along with increased apoptosis and disruption of microtubule dynamics. Furthermore, under conditions of inhibition of autophagy with Baf, apoptosis triggered by these compounds was significantly increased. ConclusionOur collective results suggest that treatment with epothilones in combination with autophagy inhibitors present a potentially more effective chemotherapeutic approach for ovarian cancer.

Follicle-Stimulating Hormone Is an Autocrine Regulator of the Ovarian Cancer Metastatic Niche Through Notch Signaling.

The association between the upregulated Notch and FSH signaling and ovarian cancer is well documented. However, their signaling has been investigated independently and only in the primary tumor tissues. The aim of this study was to investigate the interactive effects of FSH and Notch signaling on ovarian cancer proliferation, formation, and maintenance of disseminated ovarian cancer cells. The roles of Notch and FSH in ovarian cancer pathogenesis were investigated with ovarian cancer cell lines and specific antibodies against Notch and FSH receptor (FSHR). FSH upregulated Notch signaling and proliferation in ovarian cancer cells. High levels of FSH were detected in the ascites of patients with serous ovarian adenocarcinoma. Spheroids from the patients’ ascites, as well as the spheroids from ovarian cancer cell lines under low attachment culture conditions, expressed FSHβ subunit mRNA and secreted the hormone into the medium. In contrast, primary ovarian tumor tissues and cell line monolayers expressed very low levels of FSHβ. Ovarian cancer cell spheroids also exhibited higher expression of FSH receptor and Notch downstream genes than their monolayer counterparts. A combination of FSHR and Notch antagonistic antibodies significantly inhibited spheroid formation and cell proliferation in vitro. This study demonstrates that spheroids in ascites express and secrete FSH, which regulates cancer cell proliferation and spheroidogenesis through Notch signaling, suggesting that FSH is an autocrine regulator of cancer metastasis. Furthermore, Notch and FSHR are potential immunotherapeutic targets for ovarian cancer treatment.

Study on the expression and meaning of FBXW7 and ENO1 in high and low grade serous ovarian adenocarcinoma tissue

Objective To investigate the expression and significance of FBXW7 and ENO1 in ovarian serous adenocarcinoma of different grades. Methods Immunohistochemistry was used to study the expression of FBXW7 and ENO1 in 60 cases of ovarian serous adenocarcinoma.The relationship between FBXW7 and ENO1 proteins and the prognosis of ovarian serous adenocarcinoma was analyzed. Results The positive rate of FBXW7 expression was 22.5% (9/40) in 40 cases of ovarian high grade adenocarcinoma and 10 cases in 15 cases of normal oviduct.The positive rate of FBXW7 expression was 66.7% (10/15), and the difference was statistically significant (P=0.003). The expression of FBXW7 in 20 cases of low grade adenocarcinoma was 5 cases, and the positive rate was 25.0%(5/20). In 15 cases of normal ovarian tissue, 9 cases were positive, and the positive rate was 60.0%(9/15). The difference was statistically significant (P=0.04). The expression of ENO1 protein was 27 in 40 cases of high grade adenocarcinoma, and the positive rate of expression was 67.5%(27/40).5 cases were positive in 15 normal fallopian tubes, and the positive rate was 33.3%(5/15). The difference was statistically significant (P=0.024). The expression of ENO1 protein was 15 in 20 cases of low grade adenocarcinoma, and the positive rate of expression was 75.0%(15/20). In 15 cases of normal ovarian tissue, 4 cases were positive, and the positive rate was 26.7%(4/15). The difference was statistically significant (P=0.006). There was no correlation between the low expression of FBXW7 and the high expression of ENO1 in high grade ovarian adenocarcinoma (P=0.199), but there was a significant correlation between the low expression of FBXW7 and the high expression of ENO1 in low grade ovarian adenocarcinoma (P<0.05). In low grade serous adenocarcinoma, the 5-year survival rates were 44.4% and 32.1% respectively, with no significant difference (P=0.052). In ovarian high-grade serous adenocarcinoma, the 5-year survival rates of high-expression group and low-expression group were 20.0% and 7.7%, respectively, with no significant difference (P=0.097). In low grade ovarian serous adenocarcinoma, the 5-year survival rate was 7.4% in high expression group and 50.0% in low expression group (P=0.023). The 5-year survival rates of ENO1 in high-grade serous adenocarcinoma were 0% and 40.0% in high-expression group and low-expression group respectively (P=0.001). Conclusion The low expression of FBXW7 in ovarian adenocarcinoma suggests that FBXW7 may be a tumor suppressor gene in ovarian serous adenocarcinoma, and ENO1 may be an oncogene in ovarian adenocarcinoma.The high expression of FBXW7 in serous adenocarcinoma indicates that ENO1 may be an oncogene, and the survival rate of FBXW7 in serous adenocarcinoma is higher than that in low expression group.The survival rate of the high-expression group was lower than that of the low-expression group.Therefore, they may become a new diagnostic index and therapeutic target for ovarian serous adenocarcinoma. Key words: FBXW7; ENO1; Advanced ovarian serous carcinoma; Ovaroian low-level serous carcinoma; Immunohistochemistry method

Abstract B74: Identification of prognostic molecular subtypes of ovarian serous cystadenocarcinoma by isoform-level gene expression analysis

Abstract Characterization of molecular aberrations in ovarian serous cystadenocarcinoma (OVSC) is highly crucial for developing precision medicine approaches. While gene-level analysis of gene expression profiles of The Cancer Genome Atlas (TCGA) OVSC samples revealed four molecular subgroups (differentiated, proliferative, mesenchymal, and immunoreactive), the association of the derived subtypes with the clinical and pathologic features, such as stage, cytoreduction, and chemotherapy, is not statistically significant. Considering the fact that the majority of human genes produce multiple transcript-variants that could be involved in different functional pathways, we reanalyzed the TCGA exon-array expression profiles at the transcript variant or isoform level by following the novel informatics methodology we developed and successfully applied for glioblastoma (Pal et al., Nucleic Acids Res 2014). Non-negative matrix factorization (NMF) clustering of TCGA ovarian serous adenocarcinoma samples, based on isoform-level gene-expression profiles, recaptured the four known molecular subgroups but switched the subtype for 26% of the samples, resulting in significant (p=0.009) survival differences among the refined subgroups. The survival analysis was further extended to multivariate Cox proportional hazard model considering the different subgroups, and the clinical parameters as the predictor variables. Patients classified under mesenchymal group (HR = 1.44; 95% CI, 0.99-2.08; P &amp;lt; 0.05) and in stage III (HR = 2.39, 95%of CI, 1.25-4.56, P &amp;lt; 0.05), IV (HR= 3.16; 95% of CI, 1.58-6.32, P &amp;lt; 0.05) were identified to be at high risk or decreased survival, while those receiving intraperitoneal (IP) chemotherapy (HR, 0.63; 95% of CI, 0.38-1.03, P &amp;lt; 0.1) were associated with longer survival. We are in the process of translating the isoform-level gene signature into clinically adaptable molecular subtyping assay and validating on an independent patient cohort, with potential implications for developing robust diagnostic assays for OVSC patient stratification. Citation Format: Arunima Shilpi, Brandon L. Seagle, Manoj Kandpal, Shohreh Shahabi, Ramana V. Davuluri. Identification of prognostic molecular subtypes of ovarian serous cystadenocarcinoma by isoform-level gene expression analysis. [abstract]. In: Proceedings of the AACR Conference: Addressing Critical Questions in Ovarian Cancer Research and Treatment; Oct 1-4, 2017; Pittsburgh, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(15_Suppl):Abstract nr B74.

Sphincter-preserving surgery for recurrent pelvic malignancy using a hybrid procedure of open laparotomy and transanal endoscopic approach

BackgroundSurgery for the treatment of recurrent pelvic malignancy is challenging. Sphincter-preserving surgery (SPS) has been applied in limited cases. Transanal endoscopic approach (TEA) has been used for primary rectal cancer, predominantly for hybrid transabdominal-transanal total mesorectal excision. Here, we describe the use of TEA as a hybrid approach in a case of recurrent ovarian cancer.Case presentationA 59-year-old woman had recurrence of serous ovarian adenocarcinoma in the vaginal stump, near the site of anastomosis from a rectal resection 18 months previously. We used a hybrid approach comprising conventional open laparotomy and TEA to accomplish sphincter preservation. In addition to sphincter preservation, TEA allowed for the creation of a “terminal” space, which was made by anterior dissection between the rectum and the vagina. We employed TEA to create an opening in the scar tissue along the sacrum, which was used as a “guide” for pelvic dissection to prevent nerve injury. After exteriorization of the tumor, bowel continuity was achieved by hand-sewn coloanal anastomosis with a protective diverting ileostomy. Pathological examination revealed no involvement of the surgical margins. The diverting ileostomy was taken down 8 months postoperatively.ConclusionA hybrid approach comprising conventional open laparotomy and TEA allowed for safe and secure SPS and complete excision of a recurrent pelvic malignancy. This hybrid surgical approach expands the use of SPS in highly selected cases.

A novel multiple biomarker panel for the early detection of high-grade serous ovarian carcinoma

IntroductionSince the majority of patients are diagnosed at an advanced stage, ovarian cancer remains the most lethal gynecologic malignancy. There is no single biomarker with the sensitivity and specificity required for effective cancer screening; therefore, we investigated a panel of novel biomarkers for the early detection of high-grade serous ovarian carcinoma. MethodsTwelve serum biomarkers with high differential gene expression and validated antibodies were selected: IL-1Ra, IL-6, Dkk-1, uPA, E-CAD, ErbB2, SLPI, HE4, CA125, LCN2, MSLN, and OPN. They were tested using Simple Plex™, a multi-analyte immunoassay platform, in samples collected from 172 patients who were either healthy, had benign gynecologic pathologies, or had high-grade serous ovarian adenocarcinomas. The receiver operating characteristic (ROC) curve, ROC area under the curve (AUC), and standard error (SE) of the AUC were obtained. Univariate ROC analyses and multivariate ROC analyses with the combination of multiple biomarkers were performed. ResultsThe 4-marker panel consisting of CA125, HE4, E-CAD, and IL-6 had the highest ROC AUC. When evaluated for the ability to distinguish early stage ovarian cancer from a non-cancer control, not only did this 4-marker panel (AUC=0.961) performed better than CA 125 alone (AUC=0.851; P=0.0150) and HE4 alone (AUC=0.870; P=0.0220), but also performed significantly better than the 2- marker combination of CA125+HE4 (AUC=0.922; P=0.0278). The 4-marker panel had the highest average sensitivity under the region of its ROC curve corresponding to specificity ranging from 100% down to ~95%. ConclusionThe four-marker panel, CA125, HE4, E-CAD, and IL-6, shows potential in detecting serous ovarian cancer at earlier stages. Additional validation studies using the biomarker combination in ovarian cancer patients are warranted.

Overexpression of tyrosine kinase receptor B promotes metastasis of ovarian serous adenocarcinoma by lymphangiogenesis.

The aim of this study was to detect the expression features of tyrosine kinase receptor B (TrkB) and analyze the possible correlation between TrkB expression and lymph vessel density (LVD) in metastasis of ovarian serous adenocarcinoma. An immunohistochemical method was used to evaluate TrkB expression in 139 ovarian tumor sections (103 primary ovarian serous adenocarcinomas and 36 serous adenomas) and investigate the correlation between TrkB expression and LVD, which was estimated by means of VEGFR-3 assessment. TrkB was significantly upregulated in serous adenocarcinomas and absent in serous adenomas. There was no association between TrkB expression and the histological grade of cancer cells. The expression of TrkB was correlated with surgicopathological stage and metastasis in serous adenocarcinomas. The level of TrkB was higher in advanced-stage than in early-stage disease. TrkB was overexpressed in metastatic lesions compared with the corresponding primary lesions. Furthermore, a positive correlation between TrkB expression and LVD in serous adenocarcinomas was observed. TrkB was overexpressed in ovarian serous adenocarcinomas and might be involved in cancer metastasis by associated lymphangiogenesis.

Human cytomegalovirus in high grade serous ovarian cancer possible implications for patients survival.

Patients diagnosed with high grade serous ovarian adenocarcinoma have a poor prognosis. Recently human cytomegalovirus (HCMV) has been detected in several tumors. Here, we evaluated HCMV in ovarian cancer tissue specimens obtained at pre- and postchemotherapy tumor resection.Available paraffin embedded ovarian cancer tissues from matched pre- and postchemotherapy tumor resection specimens (i.e., diagnostic excisional biopsy prechemotherapy; DEBPC) and neoadjuvant chemotherapy followed by interval debulking surgery (NACT + IDS) from 10 patients with stage IIIC-IV high grade serous ovarian carcinoma (HGS) diagnosed between years 2007 and 2008 at Karolinska University Hospital were examined for HCMV immediate-early protein (HCMV-IE), tegument protein pp65, and nucleic acid (β2.7) by immunohistochemistry and in situ hybridization.HCMV-IE and pp65 were detected in 8/10 (80%), 4/9 (44%) and in 4/10 (40%), 5/8 in ovarian cancer tissue specimens from DEBPC and NACT + IDS, respectively. HCMV-β2.7 was detected in all available tissue sections obtained from DEBPC and NACT + IDS. Patients with HCMV-IE or pp65 positive cells in their ovarian tumors at IDS after NACT had a median overall survival of 23.4 and 18.2 months, respectively, compared to 29.6 and 54 months, respectively, in those who did not express HCMV proteins in their tumors.In conclusion, HCMV proteins and nucleic acids are frequently detected at different levels in HGS ovarian carcinoma. Despite the limitation of our study, shorter median overall survival of patients with HCMV-IE and pp65 in their tumor highlights the need to further investigate the role of HCMV in ovarian cancer patients.

Expressions and clinical significances of paired box gene 2 and cyclin D1 in advanced ovarian serous carcinoma

Objective: To investigate the expressions and clinical significances of paired box gene 2 (Pax2) and cyclin D1 protein in advanced ovarian serous carcinoma. Methods: From January 2003 to December 2013, the pathologic tissues of 202 patients with advanced ovarian serous cancer (Ⅲ-Ⅳ) who underwent initial cytoreductive surgery were collected. The expressions of Pax2 and cyclin D1 protein were detected by immunohistochemistry in tissue microarray. The relationships of their expressions with the clinicopathological features and prognosis of the patients were analyzed. Results: The positive rate of Pax2 protein expression of the 202 patients with ovarian serous adenocarcinoma was 24.8% (50/202) and that of cyclin D1 was 25.2% (51/202). The expressions of Pax2 and cyclin D1 were not significantly related with age, clinical stage and pathological grade of ovarian serous adenocarcinoma patients (P>0.05). The median overall survival (OS) time of Pax2-negative patients was 53 months and the progression-free survival (PFS) time was 29 months. The median OS time of Pax2-positive patients was 66 months and PFS time was 33 months, the OS of Pax2-negative patients was significant different from that of Pax2-positive patients (χ(2)=4.06, P=0.04). The median PFS time of Pax2-negative patients was not significant different from that of Pax2-positive patients (χ(2)=2.43, P=0.11). The median OS time of cyclin D1-negative patients was 62 months and PFS time was 30 months. The median OS time of cyclin D1-positive patients was 48 months and PFS time was 22 months. The median OS time of cyclin D1-negative patients was significantly different from that of cyclin D1-positive patients (χ(2)=4.71, P=0.03), while the median PFS time of cyclin D1-negative patients was marginally different from that of cyclin D1-positive patients (χ(2)=0.59, P=0.41). Multivariate analysis showed that the expression of Pax2 was an independent factor of the prognosis for patients with ovarian serous adenocarcinoma (RR=0.597, 95% CI 0.371-0.962, P<0.034). Conclusion: The expressions of Pax2 and cyclin D1 are associated with the prognosis of patients with advanced ovarian serous adenocarcinoma while Pax2 is an independent prognostic factor.

Two independent incidences of skin metastases in the umbilicus and abdominal wall in ovarian serous adenocarcinoma

Rationale:Advanced ovarian cancer is usually associated with intra-abdominal metastases and while it commonly spreads directly to the omentum, intestine, liver, or other organs, it can also metastasize through the lymphatic channels and the hematogenous pathway. With an increasing number of invasive operations being performed with chemoradiotherapy, the incidence of extra-abdominal metastases has risen. Nevertheless, ovarian cancer with skin metastases is quite rare.Patient concerns:We report a case of ovarian cancer with two independent incidences of skin metastases in the umbilicus and abdominal wall.Diagnoses:The patient was a 67-year-old woman who was diagnosed with ovarian cancer stage IIIC and underwent cytoreductive surgery. A solitary brown cauliflower-like metastatic lesion, approximately 6 × 5 × 4 cm was identified in the umbilicus area two years after primary surgery. During tumorectomy, intraoperative exploration revealed that while the tumor was located close to the peritoneum, there was no penetration.Interventions:The patient recovered well and received multiple rounds of chemotherapy. Ten months later, the patient presented with skin lesions located on the abdominal wall that grew rapidly and spread from the lower abdomen wall to the bilateral waist and femoral skin. These lesions were multiple, ulcerated, rough heliotrope plaques that produced a foul-smelling faint yellow liquid. Biopsy analysis revealed skin metastasis of poorly differentiated serous adenocarcinoma.Outcomes:The patient was treated with chemotherapy but died 3 months after the skin metastasis occurred for the second time.Lessons:Ovarian cancer with skin metastasis is a rare condition with poor prognosis. Pathological diagnosis of early skin lesions is essential for ovarian cancer patients and that systemic and local disease should be treated with surgery or palliative therapy in order to provide patients with the best chances of survival. Tumorectomy is appropriate when lesions are isolated and when the patient's performance status is good. However, systemic therapy including chemotherapy and radiotherapy should be considered when skin lesions are associated with severe intro-abdominal disease.

Ovarian serous carcinomas acquire cisplatin resistance and increased invasion through downregulation of the high-temperature-required protein A2 (HtrA2), following repeated treatment with cisplatin.

High-temperature-required protein A2 (HtrA2) is one of the serine proteases related to apoptosis. HtrA2 protein expression has been associated with cisplatin resistance and poor prognosis in ovarian serous adenocarcinoma (SAC). The aim of this study was to understand the influence of HtrA2 on repeated treatment with cisplatin. The change in HtrA2 expression in 31 ovarian cancers was investigated by immunohistochemical analysis, before and after cisplatin-based chemotherapy, and the association between HtrA2 expression after chemotherapy and prognosis was analyzed. The association between the change in HtrA2 and proteins associated with LATS1 in ovarian serous cancer cell lines after repeated treatment with cisplatin was evaluated in vitro. In immunohistochemical analysis, repeated cisplatin treatment induced downregulation of HtrA2 protein expression, before and after cisplatin-based chemotherapy in SAC. Progression-free survival and overall survival of SAC with low expression of HtrA2 were worse than those with high expression. In vitro analysis using cisplatin-sensitive ovarian cancer cell lines, KF28, and cisplatin-resistant cancer cell lines, KFr13, obtained from KF28 by repeated cisplatin treatment, showed that HtrA2 protein expression was lower in KFr13 than in KF28. Furthermore, KFr13 had a higher invasive capacity than KF28. Next, downregulation of HtrA2 transfected with an HtrA2-specific siRNA in KF28 had not only cisplatin resistance, but also more invasive capacity than those with non-specific siRNA. Repeated treatment with cisplatin downregulated the HtrA2 protein, which led to cisplatin resistance and increased invasive capacity. Thus, HtrA2 might be a biomarker of response to cisplatin treatment and prognosis, after cisplatin-based chemotherapy.

P38MAPK family isoform p38α and activating transcription factor 2 are associated with the malignant phenotypes and poor prognosis of patients with ovarian adenocarcinoma

This study was to identify the biomarkers for the malignancy and poor prognosis in patients with ovarian cancer. The protein expression of p38MAPK family isoform p38α (p38α) and activating transcription factor 2 (ATF2) was measured in 120 ovarian serous adenocarcinomas and 34 normal fallopian tubes using immunohistochemistry. Stable OV-90 cells expressing p38α and ATF2 inhibitor were established using shRNA lentivirus. Cell proliferation, invasion, and migration were analyzed in vitro. Tumor growth and chemosensitivity were investigated in xenograft tumor models. The percentage of positive p38α and ATF2 expression was significantly higher in ovarian serous adenocarcinomas than that in normal fallopian tubes. Positive p38α and ATF2 expression were significantly associated with high clinical stage (III/IV), lymph node metastasis, and shorter overall survival. Silencing of p38α and ATF2 gene expression in OV-90 cells significantly inhibited cell proliferation, migration, and invasion in vitro. OV-90 cells with p38α and ATF2 gene being silenced grew significantly slow and were significantly sensitive to the chemotherapy compared to cells with high p38α and ATF2 expression. p38α and ATF2 expression play a crucial role in the malignant phenotypes of ovarian tumor cells and are a marker for the poor prognosis of patients with ovarian serous adenocarcinomas.

Case 2 – A case of secondary platinum-resistant ovarian cancer

Patients with ovarian cancer relapsing during first-line treatment (refractory) or in the following few months (platinum-resistant) are a very heterogeneous group with various biological tumor behaviors. As this condition is linked to an unfavorable prognosis, the main objective of treatment is to palliate symptoms and preserve quality of life. While traditional chemotherapy may help to achieve this, new biological agents that have been introduced or are under development are expected to improve the quality of life and outcomes for patients with advanced ovarian cancer. This case report describes the clinical history of a 67-year-old woman with bilateral ovarian high grade serous papillary adenocarcinoma with lymph node and omentum metastasis.

Novel near-diploid ovarian cancer cell line derived from a highly aneuploid metastatic ovarian tumor.

A new ovarian near-diploid cell line, OVDM1, was derived from a highly aneuploid serous ovarian metastatic adenocarcinoma. A metastatic tumor was obtained from a 47-year-old Ashkenazi Jewish patient three years after the first surgery removed the primary tumor, both ovaries, and the remaining reproductive organs. OVDM1 was characterized by cell morphology, genotyping, tumorigenic assay, mycoplasma testing, spectral karyotyping (SKY), and molecular profiling of the whole genome by aCGH and gene expression microarray. Targeted sequencing of a panel of cancer-related genes was also performed. Hierarchical clustering of gene expression data clearly confirmed the ovarian origin of the cell line. OVDM1 has a near-diploid karyotype with a low-level aneuploidy, but samples of the original metastatic tumor were grossly aneuploid. A number of single nucleotide variations (SNVs)/mutations were detected in OVDM1 and the metastatic tumor samples. Some of them were cancer-related according to COSMIC and HGMD databases (no founder mutations in BRCA1 and BRCA2 have been found). A large number of focal copy number alterations (FCNAs) were detected, including homozygous deletions (HDs) targeting WWOX and GATA4. Progression of OVDM1 from early to late passages was accompanied by preservation of the near-diploid status, acquisition of only few additional large chromosomal rearrangements and more than 100 new small FCNAs. Most of newly acquired FCNAs seem to be related to localized but massive DNA fragmentation (chromothripsis-like rearrangements). Newly developed near-diploid OVDM1 cell line offers an opportunity to evaluate tumorigenesis pathways/events in a minor clone of metastatic ovarian adenocarcinoma as well as mechanisms of chromothripsis.

The expression profile and prognostic value of APE/Ref-1 and NPM1 in high-grade serous ovarian adenocarcinoma.

To analyze the expression trends and clinical significance of Apurinic/Apyrimidinic Endodeoxyribonuclease 1 (APE1/Ref-1) and Nucleophosmin (NPM1) proteins in high-grade serous ovarian adenocarcinoma (HGSC). The expressions of APE1/Ref-1 and NPM1 proteins in 94 patients with HGSC were determined using the immunohistochemical (IHC) method, and their relationships with clinicopathological features were analyzed by the χ2 test or Fisher's exact test. The follow-up data, Cox proportional hazards univariate and multivariate survival analyses were integrated to evaluate the prognostic factors affecting patients with HGSC. In the normal fallopian tubes, APE1/Ref-1 and NPM1 protein were mainly distributed in the nuclear. The HGSC experienced changes in the cellular localization of APE1/Ref-1 and NPM1 protein expressions, which were abnormally expressed in the cytoplasm. The rates of abnormal cytoplasmic expression of APE1/Ref-1 and NPM1 proteins in 94 patients with HGSC were 69.1% and 73.4%, respectively, which were significantly higher than the normal fallopian tube tissues (p < 0.05). The abnormal cytoplasmic APE1/Ref-1 and NPM1 are significantly correlated with the lymph node metastasis, chemosensitivity, FIGO staging, and prognosis. The COX multivariate survival analysis showed that the abnormal expression of APE1/Ref-1 protein, FIGO staging, and lymph node metastasis are independent prognostic factors. Collectively, the abnormal cytoplasmic APE1/Ref-1 and NPM1 proteins are associated with the oncogenic progression and chemoresistance of HGSC, and predict a poor prognosis.