Agnaldo Lopes da Silva Filho

Ovarian cancer cell heterogeneity and paclitaxel response in vitro 2D and 3D cancer cell models and xenograft growth in the chicken chorioallantoic membrane (CAM)

Among gynecological malignancies, epithelial ovarian cancer (EOC) is the most lethal, comprising a heterogeneous group of diseases. Paclitaxel is a standard chemotherapeutic agent and an important option for EOC treatment. However, chemotherapy often fails, contributing to a high rate of EOC recurrence. One of the models that best explains the cellular heterogeneity, disease aggressiveness, chemoresistance, recurrence risk, and metastasis observed in ovarian tumors is the presence of cancer stem cells (CSCs). This study aimed to assess the phenotypic changes in three EOC cell lines (TOV-21G, SKOV-3, and OV-90) cultured in monolayer (MN) and tumorsphere (CSCs enrichment-TS) models. After treatment with paclitaxel, we studied different cell subpopulations by immunophenotyping using CSC markers (CD44/CD24) and mesenchymal markers (CD117/CD146). The relative expression of mRNAs (CASP8, TNFRSF10B, TP53, and BCL2) and miRNAs (miR-26a, miR-125b-5p, miR-181c, miR-17-5p, and miR-221) was evaluated by qPCR. In addition, we assessed the growth capacity of residual cells treated with paclitaxel using the chorioallantoic membrane (CAM) model to study disease relapse. After paclitaxel exposure, OV-90 TS and TOV-21G TS cells showed an increase in cell growth ability, while SKOV-3 MN cells maintained colony formation capability. SKOV-3 MN cells were enriched in the CSC-like subset (CD24

The OvarianTag™ Biomarker Panel Emerges as a Prognostic Tool to Guide Clinical Decisions in Cisplatin-Based Treatment of Epithelial Ovarian Cancer

Epithelial ovarian cancer (EOC) is the most lethal gynecologic malignancy, often diagnosed at an advanced stage due to its asymptomatic progression. The high recurrence rate and development of platinum-based chemotherapy resistance contribute to its poor prognosis. Despite advancements in molecular profiling, predictive biomarkers for chemotherapy response and recurrence risk remain limited. In this study, we developed OvarianTag™, a biomarker panel integrating apoptosis and necroptosis pathways, to predict chemotherapy benefit and disease progression in EOC patients. This observational study was conducted in two phases. In the first phase, 45 patients were recruited, and RNA was extracted from fresh ovarian tissues (normal, benign, and malignant). qRT-PCR was performed to assess the relative expression of genes involved in apoptosis and necroptosis-regulated cell death pathways. Machine learning algorithms were applied to identify the relevant prognostic markers, leading to the development of OvarianTag™. In the second phase, 55 additional EOC patients were included, and their formalin-fixed, paraffin-embedded (FFPE) tumor samples were analyzed using qRT-PCR. The classifier algorithm incorporated hierarchical clustering to stratify patients based on gene expression profiles. Significant differences in TNFRSF10C/TRAIL-R3, TNFRSF10B/TRAIL-R2, and CASP8 expression levels were observed between patient groups. CASP8 downregulation was strongly correlated with platinum resistance and a poor prognosis. Decision tree models achieved 83.3% accuracy in predicting platinum response and 79.2% accuracy in recurrence risk stratification. The OvarianTag™ classifier demonstrated high sensitivity and specificity in identifying high-risk patients, supporting its potential as a prognostic tool. The OvarianTag™ panel provides a novel approach for risk stratification in EOC, integrating apoptosis and necroptosis pathways to refine chemotherapy response prediction and recurrence risk assessment. This molecular assay has the potential to guide personalized treatment strategies, enhancing clinical decision-making and improving patient outcomes. Further validation in independent cohorts is warranted to establish its clinical utility.

Exploring cervical cancer mortality in Brazil: an ecological study on socioeconomic and healthcare factors

To evaluate the correlation between socioeconomic and healthcare factors and cervical cancer mortality rates, as well as the accessibility to prevention and treatment across Brazilian states and macroregions. The aim is to highlight the multifaceted challenge of addressing cervical cancer mortality, particularly in low-and middle-income countries. This ecological study analyzed public data from the Brazilian National Institute of Cancer (INCA), the National Institute of Geography and Statistics (IBGE), and the Brazilian Ministry of Health. Data were collected on indicators such as the Human Development Index (HDI), physician density, average household income, human papillomavirus (HPV) vaccine coverage, Pap smear screening rates, radiotherapy machine density, and non-White population rates by state and macroregion across Brazil. Spearman's rank correlation test and simple linear regression analysis were employed. Cervical cancer mortality rates are statistically lower in women with health insurance, positive self-perception of health, located in states with a higher HDI, higher per capita household income, greater density of physicians, and higher availability of radiotherapy machines. In contrast, mortality rates proportionally increase according to poverty levels, as expected, and rates of non-White population. Considering public health, HDI scores significantly affected Pap smear test coverage, the number of radiotherapy machines, and HPV vaccine uptake. The North and the Southeast regions have, respectively, the lowest and the highest socioeconomic indicators, proportional to their mortality rates. No significant correlation was found between mortality rates and HPV vaccine or Pap smear coverage. Cervical cancer mortality in Brazil is significantly influenced by socioeconomic and healthcare disparities. This study provides a data-driven basis for public health strategies that address both medical and social determinants of health.

Germline Mutations Landscape in a Cohort of the State of Minas Gerais, Brazil, in Patients Who Underwent Genetic Counseling for Gynecological and Breast Cancer

Abstract Objective The present study evaluated the profile of germline mutations present in patients who underwent genetic counseling for risk assessment for breast cancer (BC), ovarian cancer (OC), and endometrial cancer (EC) with a possible hereditary pattern. Methods Medical records of 382 patients who underwent genetic counseling after signing an informed consent form were analyzed. A total of 55.76% of patients (213/382) were symptomatic (personal history of cancer), and 44.24% (169/382) were asymptomatic (absence of the disease). The variables analyzed were age, sex, place of birth, personal or family history of BC, OC, EC, as well as other types of cancer associated with hereditary syndromes. The Human Genome Variation Society (HGVS) nomenclature guidelines were used to name the variants, and their biological significance was determined by comparing 11 databases. Results We identified 53 distinct mutations: 29 pathogenic variants, 13 variants of undetermined significance (VUS), and 11 benign. The most frequent mutations were BRCA1 c.470_471delCT, BRCA1 c.4675 + 1G > T, and BRCA2 c.2T> G. Furthermore, 21 variants appear to have been described for the first time in Brazil. In addition to BRCA1/2 mutations, variants in other genes related to hereditary syndromes that predispose to gynecological cancers were found. Conclusion This study allowed a deeper understanding of the main mutations identified in families in the state of Minas Gerais and demonstrates the need to assess the family history of non-gynecological cancer for risk assessment of BC, OC, and EC. Moreover, it is an effort that contributes to population studies to evaluate the cancer risk mutation profile in Brazil.

Hormone therapy after risk-reducing surgery in patients with BRCA1/BRCA2 mutation: evaluation of potential benefits and safety

SUMMARY Women with mutations in the BRCA 1 and 2 genes are at increased risk for ovarian and breast cancer and therefore candidates for risk-reducing surgery, including salpingo-oophorectomy and mastectomy. Risk-reducing salpingo-oophorectomy (RRSO) is considered the most effective prophylactic measure for ovarian cancer prevention in this group of patients. This procedure involves loss of ovarian function and induced menopause. Estrogen therapy is the most effective treatment for controlling vasomotor symptoms and improving the quality of life of climacteric women. However, the potential hormonal stimulation of these tumors and the risk of breast cancer are a concern regarding the safety of hormone replacement therapy (HRT) in this population. This article aims to review the current evidence regarding the potential benefits and safety of HRT after RRSO.

Adnexal mass: diagnosis and management

Apoptosis-related gene expression can predict the response of ovarian cancer cell lines to treatment with recombinant human TRAIL alone or combined with cisplatin

The objectives of this study were to determine the sensitivity of ovarian cancer (OC) cell lines (TOV-21G and SKOV-3) to cisplatin and to the recombinant human TRAIL (rhTRAIL), and to evaluate the expression profile of TNFRSF10B, TNFRSF10C, TP53TG5, MDM2, BAX, BCL-2 and CASPASE-8 genes and their participation in the resistance/susceptibility mechanism of these tumor cell lines. To determine the IC50 values associated with Cisplatin and rhTRAIL, inhibition of cell growth was observed using MTT assays in two human OC cell lines (SKOV-3 and TOV-21G). The analysis of gene expression was performed using quantitative real-time polymerase chain reaction (qRT-PCR). Both cell lines had different susceptibility profiles to the tested drugs. In the SKOV-3 cell line, the IC50 values for cisplatin and for rhTRAIL were 270.83 ug/mL and 196.5 ng/mL, respectively. The same concentrations were used for TOV-21G. Different gene expression profiles were observed in each tested cell line. CASPASE-8 and TNFRSF10B expression levels could predict the response of both the cell lines to rhTRAIL alone or the response to a combination of rhTRAIL and cisplatin. In addition, we observed a relationship between BCL-2 and BAX expression that may be helpful in estimating the proliferation rate of the OC cell lines. SKOV-3 and TOV-21G respond differently to cisplatin and rhTRAIL exposure, and expression of CASPASE-8 and TNFRSF10B are good predictors of responses to these treatments.

High-Grade Transformation in Adenoid Cystic Carcinoma of the Bartholin Gland: Case Report

Abstract Introduction In the present study, we report a case of primary adenoid cystic carcinoma (ACC) of the Bartholin gland with high-grade transformation (HGT). Adenoid cystic carcinoma of the Bartholin gland is a rare tumor and HGT has only been reported in head and neck tumors. Case Report A 77-year-old woman with a non-ulcerated vulvar lesion on the topography of the right Bartholin gland. The patient was submitted to tumor resection followed by V–Y island flap and adjuvant radiotherapy. The histopathological examination revealed primary ACC of the Bartholin gland, with areas of HGT and extensive perineural invasion. The immunohistochemical study with p53 showed a diffuse and strong positive reaction in areas with HGT. After 24 months of follow-up, the patient presented distant metastases and died, despite having undergone to chemotherapy. Conclusion As far as we know, this case is the first description in the literature of HGT in ACC of the Bartholin gland, and HGT appears to be associated with tumor aggressiveness.

Call to Eliminate Cervical Cancer in the Next Decade with a Focus on Brazil

Cervical Cancer-Related Knowledge, Attitudes, Practices and Self-Screening Acceptance Among Patients, Employees, and Social Media Followers of Major Brazilian Hospital

Background Brazil has a high burden of cervical cancer, even though it is preventable, traceable and treatable. Hence, this study evaluated levels of knowledge, attitudes and practices (KAP) related to cervical cancer screening and diagnosis and acceptance of self-screening techniques among women aged 24 and greater. Methods A cross-sectional KAP survey was administered to n = 4206 women and spanned questions relating to cervical cancer, HPV, speculum, Pap test and colposcopy. Questionnaire was disseminated through a major hospital’s social media platforms, intranet and gynecologic-oncology clinics. Logistic regressions evaluated associations between sociodemographic characteristics and knowledge, attitudes, and preventative behaviors against cervical cancer. Participants indicated willingness to try DNA-HPV self-sampling and cervix self-visualization (self-colposcopy). Findings Participants were mostly white individuals (70.5%) with higher education and from social classes A and B. They demonstrated superior levels of KAP than described in the literature, with over 57.8% having answered 80+% of questions correctly. KAP scores were predicted by social class, educational attainment, race, history of premalignant cervical lesions and geographic location. About 80% and 63% would be willing to try DNA-HPV self-sampling and cervix self-visualization, respectively. Interest in self-screening was associated with adequate attitude (OR = 1.85) and inadequate practice (OR = .83). Interpretation Adequate KAP are fundamental for the successful implementation of a self-screening program. Participants were interested in methods that provide them with greater autonomy, control and practicality. Self-screening could address barriers for under-screened women such as shame, discomfort, distance from clinics and competing commitments, enabling Brazil to reach the WHO’s cervical cancer elimination goals. It could also decrease excess medical intervention in over-screened populations by promoting shared decision-making.

The importance of the quadrivalent HPV vaccine in the elimination of cervical cancer in Brazil

Association of insulin-like growth factor II mrna-binding protein 3 (IMP3) expression with prognostic and morphological factors in endometrial cancer

Endometrial cancer (EC) is a heterogeneous disease with recurrence rates ranging from 15 to 20%. The discrimination of cases with a worse prognosis aims, in part, to reduce the length of surgical staging in cases with a better prognosis. This study aimed to evaluate the association between Insulin-like growth factor II mRNA-binding protein 3 (IMP3) expression and prognostic and morphological factors in EC. This retrospective, cross-sectional, analytical study included 79 EC patients - 70 endometrioid carcinoma (EEC) and 9 serous carcinoma (SC) - and 74 benign endometrium controls. IMP3 expression was evaluated by immunohistochemistry-based TMA (Tissue Microarray), and the results were associated with morphological and prognostic factors, including claudins 3 and 4, estrogen and progesterone receptors, TP53, and KI67. IMP3 expression was significantly higher in SC compared to EEC in both extent (p<0.001) and intensity (p=0.044). It was also significantly associated with worse prognostic factors, including degree of differentiation (p=0.024, p<0.001), staging (p<0.001; p<0.001) and metastasis (p=0.002; p<0.001). IMP3 expression was also significant in extent (p=0.002) in endometrial tumors compared with controls. In addition, protein TP53 and KI67 showed significant associations in extent and intensity, respectively. IMP3 expression was associated with worse prognostic factors studied. These findings suggest that IMP3 may be a potential biomarker for EC poorer prognosis.

Efficacy of direct oral anticoagulants versus low-molecular-weight heparin for thromboprophylaxis after gynecological cancer surgery: A systematic review and meta-analysis

To evaluate the efficacy and safety of direct oral anticoagulants (DOACs) compared to enoxaparin, a low molecular weight heparin (LMWH). for postoperative thromboprophylaxis in patients undergoing gynecologic cancer surgery. This systematic review and meta-analysis followed Cochrane Handbook guidelines and PRISMA recommendations. We searched PubMed, Embase, Scopus, and CENTRAL for randomized controlled trials (RCTs) and observational studies comparing DOACs and LMWHs for postoperative VTE prophylaxis in gynecologic cancer surgeries. Outcomes included VTE incidence, major bleeding, clinically relevant non-major bleeding (CRNMB), hospital readmission, and drug adherence. The risk of bias was assessed using the Cochrane tools (RoB-2 and ROBINS-I), and statistical analyses were conducted using Review Manager 5. Five studies (1436 patients) were included: two RCTs and three observational studies. There was no significant difference in 30-day VTE incidence between DOAC and LMWH groups (OR, 0.55; 95 % CI, 0.17 to 1.77; P = 0.31). Similarly, major bleeding rates showed no difference (OR, 1.13; 95 % CI, 0.29 to 4.42; P = 0.90). DOACs significantly reduced CRNMB events compared to LMWHs (OR, 0.44; 95 % CI, 0.23 to 0.82; P = 0.01). Hospital readmissions (OR, 0.68; 95 % CI, 0.33 to 1.41; P = 0.30) and drug adherence rates (OR, 0.95; 95 % CI, 0.36 to 2.52; P = 0.29) were comparable between the groups. DOACs provide a safe and effective alternative to LMWH for postoperative thromboprophylaxis in gynecologic cancer surgeries. The significant reduction in CRNMB events suggests a potential safety advantage of DOACs.