Epigenetic status of FBXW7 gene and its role in Ovarian cancer pathogenesis.

BACKGROUND: Chromatin immunoprecipitation (ChIP) analysis revealed that the FBXW7 gene and the long non-coding RNA (LINC01588) are potential candidates in epithelial ovarian cancer (EOC) pathogenesis. However, their exact role in EOC is not yet known. Thus, the present study sheds light on the impact of the mutations/ methylation status of the FBXW7 gene. MATERIALS AND METHODS: We used public databases to assess the correlation between mutations/ methylation status and the FBXW7 expression. Furthermore, we performed Pearson's correlation analysis between the FBXW7 gene and LINC01588. We performed gene panel exome sequencing and Methylation-specific PCR (MSP) in HOSE 6-3, MCAS, OVSAHO, and eight EOC patients' samples to validate the bioinformatics results. RESULTS: The FBXW7 gene was less expressed in EOC, particularly in stages III and IV, compared to healthy tissues. Furthermore, bioinformatics analysis, gene panel exome sequencing, and MSP revealed that the FBXW7 gene is neither mutated nor methylated in EOC cell lines and tissues, suggesting alternative mechanisms for FBXW7 gene regulation. Interestingly, Pearson's correlation analysis showed an inverse, significant correlation between the FBXW7 gene and LINC01588  expression, suggesting a potential regulatory role of LINC01588. CONCLUSION: Neither mutations nor methylation is the causative mechanism for the FBXW7 downregulation in EOC, suggesting alternative means involving the lncRNA LINC01588.

Identification and validation of a novel long non-coding RNA (LINC01465) in ovarian cancer.

Epithelial Ovarian Cancer (EOC) is a heterogeneous disease usually diagnosed at advanced stages. Therefore, early detection is crucial for better survival. Despite the advances in ovarian research, mechanisms underlying EOC carcinogenesis are not elucidated. We performed chromatin immunoprecipitation sequencing to identify genes regulated by E2F5, a transcription factor involved in ovarian carcinogenesis. Results revealed several putative candidate genes (115 protein-coding genes, 20 lncRNAs, 6 pseudogenes, and 4 miRNAs). A literature review and bioinformatics analysis of these genes revealed a novel lncRNA candidate (LINC01465) in EOC. We validated LINC01465 by quantifying its expression in EOC cell lines and selected OVSAHO and SKOV3 as a model with high LINC01465 levels. We silenced LINC01465 and performed proliferation, wound healing, invasion, and drug resistance assays. Knocking-down LINC01465 resulted in reduced migration, suggesting potential involvement in EOC. Furthermore, to identify the significance of LINC01465 in chemoresistance, we assessed the LINC01465 levels in A2780 S cells treated with malformin, which revealed higher LINC01465 expression as compared to untreated A2780S cells implying the involvement of LINC01465 in cell death. Thus, this study unraveled the repertoire of E2F5 regulated candidate genes and suggested a putative role of LINC01465 in malformin-induced cell death in EOC.

Papillary follicular variant thyroid cancer in a malignant struma ovarii: a report of a rare case.

Summary: A 33-year-old female presented in 2013 with left flank pain. Ultrasound and MRI pelvis showed a complex mass 9 × 7 cm arising from the left ovary suggestive of ovarian torsion. She underwent a laparoscopic cystectomy, but the patient was lost to follow-up. Three years later, she presented with abdominal distension. Ultrasound and CT scan revealed a solid left ovarian mass with ascites and multiple peritoneal metastasis. Investigations showed elevated CA 125, CA 19-9. Ovarian malignancy was suspected. She underwent total abdominal hysterectomy and bilateral salpingo-oophorectomy on November 2016. The histopathology confirmed a well-differentiated thyroid cancer of ovarian origin with features of a papillary follicular variant without evidence of ovarian cancer and the thyroglobulin (Tg) level was elevated, more than 400 consistent with the diagnosis of malignant struma ovarii. The follow-up post-surgery showed normalization of CA 125, CA 19-9 and Tg. The patient underwent total thyroidectomy on January 2017. The histology was benign excluding thyroid cancer metastases to the ovary. She was started on thyroxine suppression, following which she received two ablation doses 131iodine (131I) each 5.3 GBq. The Tg remains slightly elevated at less than 10. 131I WBS showed no residual neck uptake and no distant avid metastasis. She was planned for molecular analysis which may indicate disease severity. We describe a case of malignant struma ovarii with widespread metastatic dissemination and a good response to surgery and 131I treatment without recurrence after 5 years of follow-up. The Tg remains slightly elevated indicating minimal stable residual disease. Learning points: Malignant struma ovarii is a rare disease; diagnosis is difficult and management is not well defined. Presentation may mimic advanced carcinoma of the ovary. Predominant sites of metastasis are adjacent pelvic structures. Thyroidectomy and 131iodine therapy should be considered. The management should be similar to that of metastatic thyroid cancer.

E2F5 Promotes the Malignancy of Ovarian Cancer Via the Regulation of Hippo and Wnt Pathways.

Background: E2F5 is a transcription factor that is overexpressed in the early stages of ovarian cancer and has been suggested as a potential biomarker for early detection. In this study, we aimed to examine the role of E2F5 in invasion and proliferation of ovarian cancer cells. Materials and Methods: We performed cell viability, colony formation, and invasion assays using ovarian cancer cells treated with siRNA to knock down the E2F5 gene. The regulatory effects of E2F5 on proteins involved in the apoptotic, Wnt, Hippo, and retinoblastoma signaling pathways were evaluated by western blotting following E2F5 repression. In addition, we analyzed data available on Gene Expression Profiling Interactive Analysis for correlations between E2F5 and YAP, ß-catenin, cyclin D1, cdk4, and caspase-9. Results: E2F5 was highly expressed in ovarian cancer cell lines and samples when compared to the nonmalignant tissues. Downregulation of E2F5 inhibited cell viability and invasion and promoted the phosphorylation of YAP, GSK-3-ß, ß-catenin, and retinoblastoma. However, cyclin D1, cdk4, and caspase-9 were downregulated when compared to control. Conclusion: Overall, E2F5 promotes ovarian carcinogenesis via the regulation of Hippo and Wnt pathways.

FAT4 silencing promotes epithelial-to-mesenchymal transition and invasion via regulation of YAP and ß-catenin activity in ovarian cancer.

BACKGROUND: The adhesion molecule, FAT4, has a tumor suppressor function with a critical role in the epithelial-to-mesenchymal-transition (EMT) and anti-malignant growth in several cancers. No study has investigated yet its role in epithelial ovarian cancer (EOC) progression. In the present study, we examined the role of FAT4 in proliferation and metastasis, and its mechanisms of interaction in these processes. METHODS: We have performed cell viability, colony formation, and invasion assays in ovarian cancer cells treated with siRNA to knockdown FAT4 gene expression. The regulatory effects of FAT4 on proteins involved in apoptotic, Wnt, Hippo, and retinoblastoma signaling pathways were evaluated by Western blotting following FAT4 repression. Also, 426 ovarian tumor samples and 88 non-tumor samples from the Gene Expression Profiling Interactive Analysis (GEPIA) database were analyzed for the expression of FAT4. Pearson's correlation was performed to determine the correlation between FAT4 and the E2F5, cyclin D1, cdk4, and caspase 9 expressions. RESULTS: Lower expression of FAT4 was observed in ovarian cancer cell lines and human samples as compared to non-malignant tissues. This down-regulation seems to enhance cell viability, invasion, and colony formation. Silencing FAT4 resulted in the upregulation of E2F5, vimentin, YAP, ß-catenin, cyclin D1, cdk4, and Bcl2, and in the downregulation of GSK-3-ß, and caspase 9 when compared to control. Furthermore, regulatory effects of FAT4 on the EMT and aggressive phenotype seem to occur through Hippo, Wnt, and cell cycle pathways. CONCLUSION: FAT4 downregulation promotes increased growth and invasion through the activation of Hippo and Wnt-ß-catenin pathways.

Prevalence of human papilloma virus in Oman: Genotypes 82 and 68 are dominating.

BACKGROUND: Persistent infection with high-risk (HR) HPV genotypes has been associated with cervical cancer, the third cancer affecting women in Oman with a crude incidence rate of 4.7 and mortality rate of 2.5 respectively. Other types of lower-risk (LR) HPV are associated with warts in both genders worldwide. OBJECTIVES: To assess the prevalence and genotype distribution of HPV and the risk factors among women with normal and abnormal cytology. METHODS: A cross sectional study conducted between September 2014 and April 2015. 258 cervical samples were obtained from women aged 18-68 years attending the Gynaecology Out-patient Clinic. HPV genotyping was performed using a multiplex real time-polymerase chain reaction (RT-PCR) assay. RESULTS: 22 different HPV genotypes were detected in 46 women (17.8%) and included 15 HR and 7 LR genotypes. Human immunodeficiency virus (HIV) patients (P = 0.052) and oral contraceptives users (P = 0.016) showed significant association with HPV infection. CONCLUSION: The most frequently observed HPV types were HR HPV 82 and LR HPV 54. These findings show that the predominant HPV genotypes in Oman are different from those seen in worldwide studies. This finding is important to determine the potential impact of preventive measures especially new vaccines to reduce the burden of cervical cancer.

Gestational Trophoblastic Disease at Sultan Qaboos University Hospital: Prevalence, Risk Factors, Histological Features, Sonographic Findings, and Outcomes.

OBJECTIVES: We sought to assess the prevalence of gestational trophoblastic diseases (GTD) among pregnant women at Sultan Qaboos University Hospital (SQUH) and compare our results with the international studies. We also sought to determine the risk factors, histological features, sonographic findings, and outcomes in women with GTD. METHODS: We conducted a retrospective cohort study of all women diagnosed with GTD and followed at SQUH between November 2007 and October 2015. We collected data on maternal demographics, risk factors, sonographic features, histological diagnosis, follow-up period, and chemotherapy treatment from the hospital information system. RESULTS: Sixty-four women with GTD were included in the study with a mean age of 31.0±7.5 years, mean gravidity 4.0, and parity 2.0. The prevalence of GTD was 0.3% (one in 386 births), and the most common risk factors were increased maternal age and multiparity. A partial hydatidiform mole was diagnosed in 54.7%, complete hydatidiform mole in 43.8%, and invasive mole in 1.6% of women. Eleven percent of women required chemotherapy. Typical ultrasound features for partial molar pregnancy were present in 54.7% of our sample, while snowstorm appearance was seen in 89.3% of those with complete mole. Negative beta-human chorionic gonadotropin was achieved 70 days after diagnosis in 41 women. CONCLUSIONS: The awareness of the risks and complications of GTD among physicians with close follow-up is paramount. There is a need to establish a national registry of GTD cases in Oman.

Effects of maternal age, parity and pre-pregnancy body mass index on the glucose challenge test and gestational diabetes mellitus.

OBJECTIVES: To study the effects of age, parity and body mass index (BMI) on the incidence of a positive 50 g glucose challenge test (OGCT) and gestational diabetes mellitus (GDM) in healthy pregnant Omani women. METHODS: A 50 g OGCT was performed on 307 healthy pregnant Omani women at 24-28 weeks of gestation. When the venous plasma glucose concentration (VPG) reached >7.8 mmol/l after 1 h, the OGCT was considered to be positive. Women with positive OGCTs had a confirmatory diagnosis of GDM, which was established by performing a 2-h 75 g oral glucose tolerance test (OGTT). When either fasting or post-2-h 75 g OGTT values were >5.5 mmol/l or >8 mmol/l, respectively, women were considered diabetic. RESULTS: This study screened 307 women and identified 83 (27.03%) OGCT-positive and 23 (7.5%) GDM-positive cases. The incidences of a positive OGCT and GDM increased significantly with increasing maternal age from 20.0% to 2.2%, respectively, in women aged ≤25 years to 37.8% and 14.7%, respectively, in women aged >35 years (p = 0.02 and p = 0.009, respectively). The incidences of a positive OGCT and GDM increased markedly with increasing pre-pregnancy BMI, from 19.8% to 3.8%, respectively, in women with BMIs ≤25 kg/m2 to 37.8% and 9.9%, respectively, in women with BMIs >25 kg/m2 (p = 0.02 and p = 0.04, respectively). CONCLUSION: Maternal age and pre-pregnancy BMI have profound effects on the incidences of a positive OGCT and GDM.

High Levels of IgA Antibodies to Helicobacter Pylori among Omani Women during Pregnancy and after Delivery.

BACKGROUND: Helicobacter pylori (H. pylori), is a common infection in pregnant women accompanied by variations in the levels of the IgM, IgA and IgG antibody isotypes. The variations of anti-H. pylori antibodies during and after pregnancy, and the extent of protection they provide to the mother and the fetus are not completely understood. OBJECTIVES: To investigate the changes of the anti-H. pylori IgM, IgA and IgG levels in healthy Omani pregnant women during pregnancy and 3 months after delivery. METHODS: Serum samples obtained from 70 Omani healthy pregnant women, with no history of autoimmune diseases, were tested for anti-H. pylori IgM, IgA and IgG in the first trimester of pregnancy and 3 months after delivery. In parallel and as a control group, sera obtained from a group of 70 healthy non-pregnant Omani women were tested. The levels of anti-H. pylori IgM, IgA and IgG were measured using standard Enzyme Linked Immunosorbent Assays (ELISAs). RESULTS: Anti-H. pylori IgA levels were found to be significantly higher during pregnancy (p=0.046) and after delivery (p=0.02) when compared to the control group. Moreover, a significant increase in the levels of anti-H. pylori IgM, IgA and IgG was detected after delivery (p=0.002) when compared to the levels during pregnancy. CONCLUSION: Pregnancy is associated with an increase in the levels of anti-H. pylori IgA antibodies. In addition, anti-H. pylori IgM, IgG and IgA antibody levels increase after delivery.

Does Cervical Screening in Young Women Aged 20-25 Years Lead to Unnecessary and Harmful Interventions?

BACKGROUND: Cervical human papillomavirus (HPV) infection among young women (20-25 years of age) is common and normally transient. There are growing concerns that referral to a colposcopy clinic may lead to unnecessary treatment with an increased risk of obstetric complications. Therefore, the purpose of this study was to determine the level of intervention for cervical abnormalities in this age group of the Northern Ireland population. MATERIALS AND METHODS: A review of all serial new patients under 25 years of age, who were referred to colposcopy clinics in Northern Ireland between January 1, 2009 to June 30, 2009 formed the basis of this study. RESULTS: During the study period, a total of 4,767 women under 25 years of age were screened. Two-hundred-and- thirty-four (4.9%) cases were referred to the colposcopy clinics. The cervical cytology results were: high-grade abnormality in 35%, and low-grade abnormality in 31% of these cases. One-hundred-and-seventy-eight (76%) of the referred women received at least one treatment. One-hundred-and-twenty-one of 234 (51.5%) women underwent an excisional treatment with histology showing the presence of high-grade abnormalities (CIN2-3) in 52%, CIN1 in 28%, and Koilocytosis or normal tissue in 20% of this sub-group of cases. CONCLUSIONS: Screening women under the age of 25 years cause unnecessary referral for colposcopy. This may also result in considerable anxiety and psychosexual morbidity. It leads to an over-treatment with a potential of negative impact on the future pregnancy outcomes (including pre-term delivery, low birth weight, and pre-term premature rupture of membranes).

Fluctuation in the Levels of Immunoglobulin M and Immunoglobulin G Antibodies for Cardiolipin and ß2-Glycoprotein among Healthy Pregnant Women.

OBJECTIVES: Antiphospholipid antibodies fluctuate during a healthy normal pregnancy. This study aimed to investigate the levels of both immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies for cardiolipin and ß2-glycoprotein (ß2GP) among healthy pregnant women. METHODS: This study was conducted between May 2010 and December 2012. A total of 75 healthy Omani pregnant women with no history of autoimmune disease were investigated during their pregnancy and 90 days after delivery at the Armed Forces Hospital in Muscat, Oman. A control group of 75 healthy Omani non-pregnant women were also investigated as a comparison. Levels of IgM and IgG antibodies for both anti-cardiolipin antibodies (ACAs) and ß2GP were measured using a standard enzyme-linked immunosorbent assay. RESULTS: The ACA IgM levels were significantly higher in the control group compared to the pregnant women (P <0.001). No significant differences were observed in the ACA IgM levels between the control group and the pregnant women after delivery. In contrast, ACA IgG levels were significantly higher during pregnancy and after delivery compared with those of the healthy control group (P = 0.007 and 0.002, respectively). The levels of ß2GP IgG were significantly higher during pregnancy than after delivery and in the control group (P = 0.001 and <0.001, respectively). CONCLUSION: In this study, ACA IgG levels increased during healthy pregnancies and after normal deliveries whereas ß2GP IgG levels increased transiently during the pregnancies. Both phenomena were found to be significantly associated with a transient decline in the levels of IgM specific for these antigens. Therefore, the levels of these antibodies may be regulated during a healthy pregnancy.

Ovarian Hernia: A rarity.

Ovarian hernias are extremely rare. The prevalence of ovaries and fallopian tubes in operable inguinal hernias is only about 2.9%. We report here an unusual case of an ovary in a hernia sac in an adult female. She presented with symptoms and signs of an incarcerated left inguinal hernia. The left ovary contained a haemorrhagic cyst and, along with the left fallopian tube and broad ligament, these were found in the sac. She underwent a left ovarian cystectomy and the inguinal hernia was repaired with mesh.

Do FIGO stage IA and small (≤2 cm) IB1 cervical adenocarcinomas have a good prognosis and warrant less radical surgery?

OBJECTIVES: There is a controversy regarding the optimal management of small cervical adenocarcinomas, and more radical surgery is often undertaken compared to similar size squamous carcinomas. We wished to determine the risk of parametrial involvement and metastatic disease and the outcome in International Federation of Gynecology and Obstetrics (FIGO) stage IA and small (≤2 cm) stage IB1 cervical adenocarcinomas. METHODS: All women with a diagnosis of International Federation of Gynecology and Obstetrics stages IA1, IA2, or IB1 cervical adenocarcinoma with a maximum tumor size of 2 cm were identified between 1999 and 2010 in Northern Ireland. A single pathologist reviewed all pathology prospectively at a cancer center tumor board. RESULTS: A total of 74 women were identified (mean age, 39 years; range, 25-72 years). In total, 36 women had stage IA1, 9 women had stage IA2, and 29 women had stage IB1 cervical adenocarcinomas. Surgical treatment ranged from local excision (cone or large loop excision of transformation zone) to radical hysterectomy and pelvic lymph node dissection; adjuvant therapy was not administered in any case. No parametrial involvement was seen in the 36 women who underwent parametrial resection. No lymph node metastasis was identified in the 45 women who underwent pelvic lymph node dissection. Lymphovascular space invasion was identified in 6 cases. No tumor recurrence or metastasis was noted during a mean follow-up of 35 months. CONCLUSIONS: The optimal management of women with IA or small IB1 cervical adenocarcinoma is controversial, and radical surgery is often undertaken. Our data suggest that there is an extremely low risk of parametrial and lymph node involvement with tumors 2 cm or smaller and a low recurrence rate. Less radical surgery may be warranted for small cervical adenocarcinomas, and this should be addressed by future studies.

Postirradiation primary vaginal angiosarcoma with widespread intra-abdominal metastasis.

We report a primary vaginal angiosarcoma with widespread intra-abdominal metastasis occurring in a 73-yr-old woman 13 yr after vaginal brachytherapy for an endometrial carcinoma. This is an extremely rare phenomenon with only 7 previously reported cases of vaginal angiosarcoma, 5 of which were associated with earlier irradiation. All of the earlier reported cases have been localized to the pelvis without metastatic disease.

Vitamin D Status in Pregnant Omanis: A disturbingly high proportion of patients with low vitamin D stores.

OBJECTIVES: The objective of this study was to determine the vitamin D status of pregnant Omanis by measurement of their circulating 25 hydroxy vitamin D levels. METHODS: Blood samples were obtained from a cohort of 103 consecutive healthy pregnant Omanis at the Armed Forces Hospital, Muscat, on their first antenatal visit. The study took place in May, June and July 2010. RESULTS: Vitamin D deficiency was present in 34 (33%) of patients (25OHD3 <25 nmol/L), 'at risk' levels were found in 67 (65%) patients (25OHD3 25-50 nmol/L); two patients (1.9%) had values between 50 and 75 nmol/L, and no patients in the optimal range >75 nmol/L. CONCLUSION: If confirmed, these findings indicate the need for vitamin D replacement during pregnancy and lactation. Although not evidence based we recommend at least 1000 IU of cholecalciferol, (vitamin D3) daily.