Common Variants in One-Carbon Metabolism Genes (MTHFR, MTR, MTHFD1) and Depression in Gynecologic Cancers.

We investigated the association between methylenetetrahydrofolate reductase (gene MTHFR 677C>T, rs1801133), 5-methyltetrahydrofolate-homocysteine methyltransferase (MTR 2756A>G, rs1805087), and methylenetetrahydrofolate dehydrogenase, cyclohydrolase and formyltetrahydrofolate synthetase 1 (gene MTHFD1 1958G>A, rs2236225)-well-studied functional variants involved in one-carbon metabolism-and gynecologic cancer risk, and the interaction between these polymorphisms and depression. A total of 200 gynecologic cancer cases and 240 healthy controls were recruited to participate in this study. Three single-nucleotide variants (SNVs) (rs1801133, rs1805087, rs2236225) were genotyped using the PCR-restriction fragment length polymorphism method. Depression was assessed in all patients using the Hamilton Depression Scale. Depression was statistically significantly more frequent in women with gynecologic cancers (69.5% vs. 34.2% in controls, p < 0.001). MTHFD1 rs2236225 was associated with an increased risk of gynecologic cancers (in dominant OR = 1.53, p = 0.033, and in log-additive models OR = 1.37, p = 0.024). Moreover, an association was found between depression risk and MTHFR rs1801133 genotypes in the controls but not in women with gynecologic cancers (in codominant model CC vs. TT: OR = 3.39, 95%: 1.49-7.74, p = 0.011). Cancers of the female reproductive system are associated with the occurrence of depression, and ovarian cancer may be associated with the rs2236225 variant of the MTHFD1 gene. In addition, in healthy aging women in the Polish population, the rs1801133 variant of the MTHFR gene is associated with depression.

Using a Modified Polysaccharide as a Hemostatic Agent Results in Less Reduction of the Ovarian Reserve after Laparoscopic Surgery of Ovarian Tumors-Prospective Study.

Background and Objectives: The study investigated whether the method of achieving hemostasis affects the ovarian reserve in patients undergoing laparoscopic surgery due to ovarian tumors or cysts. Materials and Methods: Patients with unilateral tumors or ovarian cysts, who qualified for laparoscopic tumor enucleation, were randomly selected to receive modified polysaccharides or bipolar coagulation. Ovarian reserve was analyzed by anti-Mullerian hormone (AMH) level. Results: The study included 38 patients: 19 patients in the modified polysaccharide group and 19 in the bipolar coagulation group. Patients after bipolar coagulation treatment had statistically significantly lower AMH 6 months after surgery compared to the group treated with modified starch. The levels of AMH in the study and control groups were 3.96 +/- 2.12 vs. 2.51 +/- 1.39 ng/mL, respectively; p = 0.018. A statistically significant decrease in AMH was also demonstrated in the bipolar coagulation group as compared to the preoperative assessment (p = 0.049). There was no statistically significant decrease in AMH in the group of patients treated with the modified starch. Conclusions: Using a modified polysaccharide during laparoscopic cystectomy is effective and has a positive effect on the ovarian reserve compared to the use of bipolar coagulation. Both the AMH level 6 months after surgery and the percentage decrease in AMH were more favorable in the group of patients treated with modified starch.

Altered Expression of ESR1, ESR2, PELP1 and c-SRC Genes Is Associated with Ovarian Cancer Manifestation.

Ovarian cancer remains the leading cause of death due to gynecologic malignancy. Estrogen-related pathways genes, such as estrogen receptors (ESR1 and ESR2) and their coregulators, proline-, glutamic acid-, and leucine-rich protein 1 (PELP1), and proto-oncogene tyrosine-protein kinase c-Src (SRC) are involved in ovarian cancer induction and development, still they require in-depth study. In our study, tissue samples were obtained from 52 females of Caucasian descent (control group without cancerous evidence (n = 27), including noncancerous benign changes (n = 15), and the ovarian carcinoma (n = 25)). Using quantitative analyses, we investigated ESRs, PELP1, and SRC mRNA expression association with ovarian tumorigenesis. Proteins' presence and their location were determined by Western blot and immunohistochemistry. Results showed that PELP1 and SRC expression levels were found to differ in tissues of different sample types. The expression patterns were complex and differed in the case of ovarian cancer patients compared to controls. The most robust protein immunoreactivity was observed for PELP1 and the weakest for ESR1. The expression patterns of analyzed genes represent a potentially interesting target in ovarian cancer biology, especially PELP1. This study suggests that specific estrogen-mediated functions in the ovary and ovary-derived cancer might result from different local interactions of estrogen with their receptors and coregulators.

Ovarian steroid cell tumor as an example of severe hyperandrogenism in 45-year-old woman.

Approximately, 5% of ovarian tumors have hormonal activity. Steroid cell tumors (SCTs) represent about 0.1% of all ovarian tumors. They cause hyperandrogenism associated with typical virilization. In this case report, we present 45-year-old women with unmalignant ovarian SCT-producing androgens which cause severe virilization and secondary amenorrhea lasting two years. Transvaginal ultrasound, computed tomography of adrenal glands, magnetic resonance imaging of small pelvis, laboratory tests (including serum concentration of FSH, LH, testosterone (T), androstenedione (A), dehydroepiandrosterone sulfate (DHEA-S), as well as ROMA index) were performed. During hormonal evaluation, elevated concentrations of serum T - on admission 1.72 ng/ml and one month later 3.75 ng/ml (normal range 0.08-0.82 ng/ml) and A - 24.90 ng/ml (normal range 0.40-3.40 ng/ml) were found. The ROMA index was within the normal range. Enlargement of the left ovary by solid mass 56 × 43 mm was found during ultrasound examination. Based on small pelvis MRI scan and hormonal finding, patient was qualified for laparotomy. During this procedure, the left salpingo-oophorectomy with removal of the tumor was performed. The histopathological examination identified SCT. During follow-up examination, one day after surgery, we found serum testosterone levels within normal ranges - 0.74 ng/ml (normal range 0.08-0.82 ng/ml). This case shows that hormone-producing ovarian tumors are rare but very important clinical causes of severe hyperandrogenism.

Performance of Selected Models for Predicting Malignancy in Ovarian Tumors in Relation to the Degree of Diagnostic Uncertainty by Subjective Assessment With Ultrasound.

OBJECTIVES: The study's main aim was to evaluate the relationship between the performance of predictive models for differential diagnoses of ovarian tumors and levels of diagnostic confidence in subjective assessment (SA) with ultrasound. The second aim was to identify the parameters that differentiate between malignant and benign tumors among tumors initially diagnosed as uncertain by SA. METHODS: The study included 250 (55%) benign ovarian masses and 201 (45%) malignant tumors. According to ultrasound findings, the tumors were divided into 6 groups: certainly benign, probably benign, uncertain but benign, uncertain but malignant, probably malignant, and certainly malignant. The performance of the risk of malignancy index, International Ovarian Tumor Analysis assessment of different neoplasias in the adnexa model, and International Ovarian Tumor Analysis logistic regression model 2 was analyzed in subgroups as follows: SA-certain tumors (including certainly benign and certainly malignant) versus SA-probable tumors (probably benign and probably malignant) versus SA-uncertain tumors (uncertain but benign and uncertain but malignant). RESULTS: We found a progressive decrease in the performance of all models in association with the increased uncertainty in SA. The areas under the receiver operating characteristic curve for the risk of malignancy index, logistic regression model 2, and assessment of different neoplasias in the adnexa model decreased between the SA-certain and SA-uncertain groups by 20%, 28%, and 20%, respectively. The presence of solid parts and a high color score were the discriminatory features between uncertain but benign and uncertain but malignant tumors. CONCLUSIONS: Studies are needed that focus on the subgroup of ovarian tumors that are difficult to classify by SA. In cases of uncertain tumors by SA, the presence of solid components or a high color score should prompt a gynecologic oncology clinic referral.

The Epithelial-Mesenchymal Transition Initiated by Malignant Ascites Underlies the Transmesothelial Invasion of Ovarian Cancer Cells.

The role of the epithelial-mesenchymal transition (EMT) in ovarian cancer cell progression is unquestioned. In this report, we describe that malignant ascites, fluid that accumulates in the peritoneal cavity in a large group of patients with ovarian cancer, stimulate EMT in two representative ovarian cancer cell lines (A2780, SKOV-3). In addition, we identify the ascites-derived mediators of EMT and signaling pathways initiated in the cancer cells that underlie this phenomenon. Finally, we demonstrate that EMT induced in the cancer cells in response to the malignant ascites contributes to their increased transmesothelial invasion. Altogether, our study provides new insight into the mechanistic aspects of the malignant ascites-dependent exacerbation of the intraperitoneal progression of ovarian cancer.

Correlation between allopregnanolone levels and depressive symptoms during late menopausal transition and early postmenopause.

OBJECTIVES: This observational, cross-sectional study included 140 women with climacteric symptoms. The aim of the study was to evaluate the correlation between the presence and severity of depressive symptoms and allopregnanolone levels in women during late menopausal transition and early postmenopause. METHODS: The study group was divided into two groups: 45 women in late menopausal transition and 95 early postmenopausal women. We evaluated Kupperman index, Hamilton scale and serum follicle-stimulating hormone, luteinizing hormone, 17ß-estradiol, prolactin, total testosterone, dehydroepiandrosterone sulfate and allopregnanolone levels. RESULTS: We found that serum allopregnanolone concentration was lower in early postmenopausal women compared to women in late menopausal transition; that there was a correlation between serum allopregnanolone levels in early postmenopausal women and time since last menstruation, intensity of climacteric symptoms, and intensity of depression symptoms and that there was a correlation between serum allopregnanolone levels and several depression symptoms presence (shallow sleep and symptoms of the digestive tract in women during late menopause transition; feelings of guilt, sleep disorders and general somatic symptoms in early postmenopausal women). CONCLUSION: We concluded that reproductive aging is characterized by a reduction of allopregnanolone circulating levels that correlate to Hamilton depression index in early postmenopause and presence of specific depressive symptoms during late menopausal transition and early postmenopause.

The association of CCAT2 rs6983267 SNP with MYC expression and progression of uterine cervical cancer in the Polish population.

PURPOSE: Previous studies have reported a significant contribution of NC_000008.10:g.128413305 G>T (rs6983267) single-nucleotide polymorphism (SNP) in the MYC enhancer region to the susceptibility of various cancers. However, the role of rs6983267 SNP in cervical cancer (CC) development and progression has not been demonstrated to date. Therefore, we evaluated the role of rs6983267 SNP in MYC expression in cervical cancers and non-cancerous cervical tissues. In addition, we assessed the role of this SNP in the development and progression of CC. METHODS: Using high-resolution melting analysis, we evaluated rs6983267 SNP frequency in women diagnosed with cervical squamous cell carcinoma (SCC) (n = 481) and controls (n = 502) in a Polish Caucasian population. Logistic regression analysis was employed to adjust for the effects of age, parity, oral contraceptive use, tobacco smoking, and menopausal status. RESULTS: Dividing patients based on clinical characteristics demonstrated an association of the rs6983267 genotype with tumor stage III and grade of differentiation G2 and G3. The p trend value calculated for the rs6983267 SNP in patients with stage III was 0.0006. We also observed a significant contribution of rs6983267 SNP to tumor grade of differentiation G2 and G3. Additional contributors were oral contraceptive use, smoking, and postmenopausal age. We found statistically significant increase of MYC transcript levels in cervical SCC tissues from carriers of the GG vs. T/T (p < 0.00001), G/T vs. T/T (p = 0.0002), and in the non-cancerous cervical tissues from carriers of the GG vs. T/T (p = 0.00046). CONCLUSION: The rs6983267 SNP may contribute to the increased MYC expression as well as the spread and rapid growth of cervical SCC as compared to lower grade carcinomas.

Severe hyperandrogenemia in postmenopausal woman as a presentation of ovarian hyperthecosis. Case report and mini review of the literature.

Ovarian hyperthecosis (OH) is characterized by the presence of abundant luteinized theca cells in ovaries that secret androgen. It typically presents as severe hyperandrogenism and/or virilization in postmenopausal woman. Here we describe a 66-year old woman with presentation of severe hirsutism, alopecia, clitoromegaly and laboratory finding of significantly elevated serum total testosterone concentration and hyperinsulinemia. Performed imaging studies revealed normal sized, homogeneous ovaries, signs of endometrial hypertrophy and normal adrenal glands. Due to severe hyperandrogenemia and signs of endometrial hypertrophy, the total abdominal hysterectomy with bilateral salpingo-oophorectomy has been performed. Pathological examination revealed OH and endometrial hyperplasia. Androgenic activity of ovarian stromal cells has been confirmed using alpha-inhibin histochemical staining. Postmenopausal hyperandrogenemia is a diagnostic and therapeutic challenge and the imaging studies often may be misleading and require careful and critical consideration.

The associations between serum VEGF, bFGF and endoglin levels with microvessel density and expression of proangiogenic factors in malignant and benign ovarian tumors.

AIM OF THE STUDY: To investigate whether serum levels of VEGF, bFGF and endoglin correlate with tumor VEGF and bFGF expression or microvessel density (MVD) in ovarian cancer. PATIENTS AND METHODS: Forty five patients with epithelial ovarian cancers (EOCs) and 38 patients with benign ovarian tumors (BOTs) were included into the study. Serum levels of VEGF, bFGF and endoglin were assessed using ELISA. The expression of VEGF and bFGF in tumor samples were evaluated using ELISA of supernatants obtained from tumor homogenization. MVD was analyzed using immunohistochemistry with antibodies against CD31, CD34 and CD105. RESULTS: Serum VEGF levels were significantly higher in EOCs than in BOTs (436.6pg/ml [19.67-2860] vs 295.5pg/ml [123-539], P=0.025). Serum endoglin levels were lowered in the group EOCs when compared to BOTs (33,720g/ml [12,220-73,940] vs 42,390pg/ml [19,380-56,910], P=0.015). There were no differences in bFGF levels between studied groups. EOCs have significantly higher CD105 MVD (25 vessels/mm2 [0-57] vs 6 vessels/mm2 [0-70], P<0.001) and tumor VEGF (405.9pg/mg protein [0-3000] vs 2.225 [0-634.7], P<0.001) expression than BOTs, while, bFGF expression was higher in BOTs than in EOCs (2076pg/mg protein [668.1-8718] vs 847.3pg/mg protein [188.9-8333], P=0.003). In patients with EOCs we have observed negative correlation between serum VEGF concentration and its tissue expression (r Spearman=-0.571, P=0.0261), and serum VEGF concentration correlated positively with CD34-MVD (r Spearman=0.545, P=0.0289). In a multiple regression analysis we have observed only the negative correlation between serum VEGF and CD105-MVD (r=-0.5288, P=0.0427). CONCLUSIONS: Serum VEGF is a useful marker for prediction of ovarian cancer MVD and tumor VEGF expression.

External validation of the IOTA ADNEX model performed by two independent gynecologic centers.

OBJECTIVES: The external, two-center validation of the IOTA ADNEX model for differential diagnosis of adnexal tumors. METHODS: A total of 204 patients with adnexal masses (134 benign and 70 malignant) treated at the Division of Gynecologic Surgery, Poznan University of Medical Sciences, Poland (Center I), and 123 patients (89 benign and 34 malignant) from the Department of Obstetrics and Gynecology, Clinica Universidad de Navarra, University of Navarra School of Medicine, Pamplona, Spain (Center II), were enrolled into the study. RESULTS: ADNEX achieved high accuracy in discriminating between malignant and benign ovarian tumors in both centers (79.9% and 81.3% in Centers I and II, respectively). Multiclass accuracy was substantially lower than in binary classification (malignant vs. benign): 64.2% and 74.0% in Centers I and II, respectively. Sensitivity and specificity for the diagnosis of specific tumor types in Center I were as follows: benign tumors - 72.4% and 94.3%; borderline tumors - 33.3% and 87.0%, stage I ovarian cancers - 00.0% and 91.8%; stage II-IV ovarian cancers - 68.2% and 83.1%; and metastatic tumors - 00.0% and 99.5%. Sensitivity and specificity in Center II were as follows: benign tumors - 75.3% and 97.1%; borderline tumors - 50.0% and 88.2%, stage I ovarian cancers - 40.0% and 97.5%; stage II-IV ovarian cancers - 95.0% and 88.3%; and metastatic tumors - 20.0% and 98.3%. CONCLUSIONS: ADNEX is characterized by very high accuracy in differentiating between malignant and benign adnexal tumors. However, prediction of ovarian tumor types could be more accurate.

Recurrent endometrial hyperplasia as a presentation of estrogen-secreting thecoma - case report and minireview of the literature.

Thecoma is a rare ovarian tumor, presenting usually in postmenopausal women as unilateral, benign, solid lesion. About 15% of affected patients develop endometrial hyperplasia (EH) and 20% are diagnosed with endometrial cancer. In this case report, we present 60-year-old women admitted because of recurrent spotting of 5 years duration, which started 1 year after menopause. In history, the patient underwent three times curettage procedures and once (1 year before admission) had estradiol levels typical for reproductive-age women. At admission, we found elevated serum levels of estradiol (222.5 pg/ml) and a small mass in the right ovary. The markers of germ cell tumors were negative. After the initial diagnosis, the patient was qualified for total abdominal hysterectomy with bilateral salpingo-oophorectomy. The histopathological examination and immunohistochemical staining confirmed the thecoma diagnosis. In follow-up examination after 8 weeks, we found decreased serum estradiol levels and relief of the symptoms. In conclusion, we want to underline that in cases of EH, especially in patients with a history of recurrences, the special attention should be paid for differential diagnosis. In such cases, the estrogen-secreting tumors should be excluded.

Ultrasonographic features and CA125 levels of hormonally active ovarian tumors.

OBJECTIVES: Subjective ultrasonographic assessment is currently considered to be the best method of differentiation between various types of ovarian tumors. The aim of the study was to evaluate selected ultrasonographic features and CA125 levels of hormonally active ovarian tumors. MATERIAL AND METHODS: A total of 1135 women with ovarian tumors were diagnosed between 2006 and 2014 at the Division of Gynecologic Surgery, Poznan University of Medical Sciences. Within these tumors, there were 60 hormone-secreting ovarian tumors, including: 20 granulosa cell tumors, 28 fibrothecomas, 10 dysgerminomas, 2 struma ovarii, and 9 metastatic ovarian tumors. The tumors were evaluated by ultrasonography according to the International Ovarian Tumor Analysis group criteria. Additionally, we evaluated serum CA125 levels in all patients. RESULTS: Granulosa cell tumors occurred most frequently as large unilocular-solid cysts, moderately to highly vascularized, with low-resistance vascularization. Dysgerminomas were predominantly large unilocular-solid cysts or purely solid tumors, with minimal to moderate low-resistance vascularization. Fibrothecomas were solid masses with minimal, high-resistance vascularization. Struma ovarii occurred as small, solid masses with abundant, highresistance vascularization. Metastatic ovarian tumors presented mainly as multilocular-solid tumors with strong, low-resistance vascularization. Papillary projections were most frequently observed in metastatic tumors and granulosa cell tumors in 56% and 50% of the cases respectively, although only half of granulosa cell tumors papillary projections exceeded 3 mm. Elevated CA125 levels were found only in metastatic ovarian tumors. CONCLUSIONS: Hormonally active ovarian tumors present several ultrasonographic features which may facilitate preoperative diagnosis.

The role of chemokine and non-chemokine factors in pain-related complaints in patients with endometrial cysts.

OBJECTIVES: Endometriosis is a chronic disease manifested as peritoneal endometrial implants and adhesions, as well as endometrial cysts, with pain as the dominant component. THE AIM: The aim of the study was to evaluate the role of chemokine (MCP-1, MCP-2, MIP-1, MIP-1 and RANTES) and non-chemokine (urocortin, ghrelin, and leptin) factors in the intensity of pelvic pain in women with endometrial cysts. MATERIAL AND METHODS: A total of 86 women, aged 18-38, treated laparoscopically between September 2009 and November 2012, due to adnexal changes, i.e. endometrial cysts and mature teratomas, were recruited for the study. On a numeric rating scale, i.e. PSS (Pain Sensation Scale--according to Johnson), the patients rated pain intensity. The level of pain-related stress was investigated with the Pain Distress Scale (PDS, according to Johnson). RESULTS: Medians for MCP-1, MCP-2, MIP-1, MIP-1 and RANTES concentrations were not statistically significantly different. The respondents rated pain intensity during menses as 6 and 3 points in the E and T groups, respectively (6 vs. 3 points; p=0.001). Statistically significant differences were also observed for pain intensity during work (apart from menses), (2 vs. 2 points, p=0,014) and during sexual intercourse (apart from menses) (3 vs. 1 points, p=0.006). Pain-related stress levels were higher in the T group as compared to the E group (3 vs. 5 points; p=0.007). CONCLUSIONS: It seems safe to conclude that chemokines and leptin may play a significant role in the occurrence of pain complaints among women with endometrial cysts. Further research might result in implementation of new treatment methods for pain management, especially in terms of pharmacotherapy.

Overweight, obesity and female sexuality in perimenopause: a preliminary report.

INTRODUCTION: The research was conducted among patients of the Department of Perinatology and Gynaecology of the Poznan University of Medical Sciences. Its aim was to investigate the influence of overweight and obesity on female sexuality during the perimenopausal period. Preliminary results of the research are presented in the thesis, which was as a matter of fact intended as a preliminary report. The examination of sexual functions of the patients was performed with the use of the Female Sexual Function Index (FSFI) form. MATERIAL AND METHODS: Sixty-one women during the perimenopausal period filled out the survey, with the average age of these women being 51 years. Forty-two of the examined women had an appropriate body mass index (BMI), i.e. between 18.5 and 25, while for 19 of the women, the BMI was above normal. For statistical analysis and in order to assess the differences between the two above-mentioned groups of patients, the nonparametric Mann-Whitney test was applied. A statistically significant value was assumed at p < 0.05. The results of the conducted research indicated no such difference between the women with differing BMI for the specific domains of the FSFI test. RESULTS: The results obtained show that research in the area needs to be continued. CONCLUSIONS: All the hitherto existing scientific studies also seem to indicate that the influence of overweight and obesity on female sexuality during the perimenopause has not yet been unambiguously proven. Beyond any doubt, however, sexual disorders appear in women at this time of life and the factors which determine them can vary greatly. Given the character of the situation, women ought to be supported both by a team of specialists representing different branches of medicine as well as by their relatives. The whole situation also calls for more research of the important subject matter.

Menopausal status strongly influences the utility of predictive models in differential diagnosis of ovarian tumors: an external validation of selected diagnostic tools.

OBJECTIVES: The aim of this study was to externally validate the diagnostic performance of the International Ovarian Tumor Analysis logistic regression models (LR1 and LR2, 2005) and other popular prognostic models including the Timmerman logistic regression model (1999), the Alcazar model (2003), the risk of malignancy index (RMI, 1990), and the risk of malignancy algorithm (ROMA, 2009). We compared these models to subjective ultrasonographic assessment performed by an experienced ultrasonography specialist, and with our previously developed scales: the sonomorphologic index and the vascularization index. Furthermore, we evaluated diagnostic tests with regard to the menopausal status of patients. MATERIALS AND METHODS: This study included 268 patients with adnexal masses; 167 patients with benign ovarian tumors and 101 patients with malignant ovarian tumors were enrolled. All tumors were evaluated by using trans- vaginal ultrasonography according to the diagnostic criteria of the analyzed models. MATERIALS AND METHODS: This study included 268 patients with adnexal masses; 167 patients with benign ovarian tumors and 101 patients with malignant ovarian tumors were enrolled. All tumors were evaluated by using trans- vaginal ultrasonography according to the diagnostic criteria of the analyzed models. RESULTS: The subjective ultrasonographic assessment and all of the studied predictive models achieved similar diagnostic performance in the whole study population. However significant differences were observed when pre- and postmenopausal patients were analyzed separately In the subgroup of premenopausal patients, the highest area under the curve (AUC) was achieved by subjective ultrasonographic assessment (0.931), the Alcazar model (0.912), and LR1 (0.909). Alternatively in the group of postmenopausal patients, the highest AUC was noted for the Timmerman model (0.973), ROMA (0.951), and RMI (0.938). CONCLUSIONS: Menopausal status is a key factor that affects the utility of prognostic models for differential diagno sis of ovarian tumors. Diagnostic models of ovarian tumors are reasonable tools for predicting tumor malignancy

DNMT1, DNMT3A and DNMT3B gene variants in relation to ovarian cancer risk in the Polish population.

Studies have demonstrated that changes in DNA methylation of cancer related genes can be an elementary process accounting for ovarian tumorigenesis. Therefore, we evaluated the possible association of single nucleotide polymorphisms (SNPs) of DNA methyltransferases (DNMTs) genes, including DNMT1, DNMT3B, and DNMT3A, with ovarian cancer development in the Polish population. Using PCR-RFLP and HRM analyses, we studied the prevalence of the DNMT1 rs8101626, rs2228611 and rs759920, DNMT3A rs2289195, 7590760, rs13401241, rs749131 and rs1550117, and DNMT3B rs1569686, rs2424913 and rs2424932 SNPs in patients with ovarian cancer (n=159) and controls (n=180). The lowest p values of the trend test were observed for the DNMT1 rs2228611 and rs759920 SNPs in patients with ovarian cancer (p trend=0.0118 and p trend=0.0173, respectively). Moreover, we observed, in the recessive inheritance model, that the DNMT1 rs2228611 and rs759920 SNPs are associated with an increased risk of ovarian cancer development [OR 1.836 (1.143-2.949), p=0.0114, p corr=0.0342, and OR 1.932 (1.185-3.152), p=0.0078, p cor=0.0234, respectively]. However, none of other nine studied SNPs displayed significant contribution to the development of ovarian cancer. Furthermore, haplotype and multifactor dimensionality reduction analysis of the studied DNMT1, DNMT3B, and DNMT3A polymorphisms did not reveal either SNP combinations or gene interactions to be associated with the risk of ovarian cancer development. Our results may suggest that DNMT1 variants may be risk factors of ovarian cancer.

Role of osteopontin in differential diagnosis of ovarian tumors.

AIM: The aim of this study was to evaluate the role of the serum osteopontin (OPN) level as a biomarker for discriminating between malignant and benign ovarian tumors. Furthermore, comparisons with the diagnostic usefulness of the other tests were performed. METHODS: The study included 114 consecutive women with ovarian tumors (82 benign and 32 malignant) who were referred to our division. RESULTS: A cut-off level of 28.0 ng/mL for OPN showed a sensitivity of 71.87% and a specificity of 89.02%. The area under the receiver-operator curve (ROC) was 0.812. There were no differences in diagnostic utility between OPN and the other studied tests. OPN levels were lower in patients with endometriotic ovarian cysts than in those with other benign ovarian tumors (14.00 vs 19.50 ng/mL; P = 0.018). The difference between the median OPN level in patients with endometriotic cysts (14.0 ng/mL) and those with malignant tumors (40.85 ng/mL) was also statistically significant (P < 0.0001). The calculated OPN/CA-125 ratio was significantly different between patients with endometriotic cysts (median, 0.36; range, 0.05-2.89) and those with other benign tumors (median, 1.25; range, 0.05-5.70) (P = 0.0002). There was also a statistically significant difference in the median OPN/CA-125 ratio between patients with endometrial cysts (median, 0.36; range, 0.05-2.89) and those with malignant tumors (median, 0.12; range, 0.01-3.39) (P = 0.004). CONCLUSION: The diagnostic utility of OPN is similar to that of ultrasonographic evaluation and CA-125 level assessment. Thus, OPN may be useful in differential diagnosis for less experienced ultrasonographers and is especially valuable for differential diagnosis of endometriotic cysts.

Usefulness of the HE4 biomarker as a second-line test in the assessment of suspicious ovarian tumors.

PURPOSE: The aim of our study was the evaluation of HE4 usefulness as a test in assessment of ovarian tumors which are suspicious and difficult to classify correctly via subjective ultrasound examination. METHODS: In this retrospective cohort study 253 women diagnosed with adnexal masses were examined preoperatively. Suspicious tumors (n = 145) were divided into groups of: "probably benign" (n = 70), "uncertain" (n = 34), and "probably malignant" (n = 41). "Uncertain" tumors were also assessed as "benign" (n = 11) or "malignant" (n = 23). The logistic regression model was performed to analyze if the serum marker improves the prediction of a malignant finding and net reclassification improvement (NRI) was calculated to measure diagnostic improvement. RESULTS: Within the analyzed group 85 (58.6%) benign and 60 (41.4%) malignant tumors were confirmed histopathologically. The comparison of HE4 with subjective ultrasound assessment showed lowered NRI in the entire analyzed group as well as in the groups of tumors classified as "probably benign" or "probably malignant" (NRI = -0.16; P = 0.0139 and NRI = -0.133; P = 0.0489, respectively). The analysis of logistic regression model confirmed that biomarkers do not improve diagnostic accuracy. The difference between areas under ROC for HE4 (0.891) and CA125 (0.902) was not statistically significant (P = 0.760). CONCLUSIONS: After subjective ultrasound assessment, the addition of the second-line test-HE4 as well as CA125 serum level does not improve diagnostic performance. However, HE4 evaluation satisfies the clinical expectations of diagnostic tools for ovarian tumors and, thus, may be useful to less experienced sonographers.

Analysis of false negative results of subjective ultrasonography assessment of adnexal masses.

OBJECTIVES: Currently transvaginal ultrasonography is the most effective method for ovarian tumor malignancy prediction. The aim of the study was to estimate the risk of false negative results in subjective interpretation of the ultrasound examination of ovarian tumors according to menopausal status. MATERIAL AND METHODS: 318 women (210 pre and 108 post menopause) with adnexal masses were diagnosed preoperatively between 2004 and 2010. Subjective assessment of tumor characteristics was conducted by experienced ultrasound examiner. Tumors were divided into groups of: "certainly benign" (n = 84), "probably benign" (n = 116), "uncertain" (n = 61), "probably malignant" (n = 47), and "certainly malignant" (n = 10). The percentage of false negative results was calculated among the first two groups according to menopausal status. RESULTS: There were 91 malignant and 227 benign adnexal masses diagnosed in histopathological evaluation. There was one false negative result of subjective interpretation of ultrasound findings in postmenopausal women - 1.6% (1/64). She was a 63-year-old woman with bilateral solid ovaries sized: 4.2 x 3.1 cm and 4.6 x 2.5 cm in ultrasound evaluation, where serous adenocarcinoma was diagnosed. There were three false negative results in premenopausal women - 2.2% (3/136). The first was a 34-year-old woman with a cyst with the appearance of ground glass of 19 x 11 cm in size where endometrioid ovarian adenocarcinoma was diagnosed. The second woman was a 32-year-old with a bilocular cyst 8 x 4.5 cm diagnosed with borderline mucinous tumor. The third patient was a 21-year-old woman with unilocular-solid cyst 4.2 x 3.2 cm where histopathological examination revealed borderline serous tumor. CONCLUSIONS: Subjective ultrasound evaluation of adnexal masses has high specificity but even in the group of tumors considered benign in premenopausal as well as postmenopausal women malignancy can be found. This occurs slightly more often before menopause.