Avaliação dos efeitos biológicos dos inibidores do mTOR associados ou não à radioterapia em linhagem celular de carcinoma do colo do útero

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Ciências da Saúde, Programa de Pós-Graduação em Ciências da Saúde, 2015.A via de sinalização PI3K/AKT/mTOR está frequentemente ativada nos carcinomas de células escamosas HPV-positivos, nos quais se inclui o câncer escamoso do colo do útero. Assim, esse trabalho propõe-se avaliar os efeitos biológicos dos inibidores do mTOR associados ou não à radioterapia em células de câncer do colo do útero HPV positiva. Foram utilizadas linhagens de células de câncer do colo do útero (HeLa) e queratinócito humano (HaCaT), usada como controle. Os inibidores de mTOR utilizados foram o temsirolimus, o everolimo, o resveratrol, a curcumina e o EGCG. O ensaio de viabilidade celular com o MTT foi realizado para obter a curva dose-resposta e determinação do IC50 para cada inibidor do mTOR. Para efeito de comparação entre os medicamentos, realizou-se análise da viabilidade celular em concentrações fixas de 50µM para os inibidores derivados da dieta e de 10µM para o everolimo e o temsirolimus. O pré-tratamento das células com os inibidores do mTOR no IC50 seguido por radioterapia foi realizado para avaliar efeito de radiossensibilização. O teste de lesão em monocamada foi realizado pós-tratamento com a curcumina e o everolimo. O perfil de morte celular induzido pelo tratamento com o temsirolimus, o resveratrol e a curcumina na linhagem HeLa foi avaliado pelo ensaio de citometria de fluxo. Os resultados mostraram que os inibidores do mTOR rapanálogos na concentração de 10µM e os derivados da dieta na concentração de 50µM induziram citoxicidade leve ou moderada nas 2 linhagens. Quando associados à radioterapia, os inibidores do mTOR apresentaram maior citotoxicidade em relação à radioterapia isolada. A curcumina interferiu no fechamento das feridas com as 2 linhagens de forma mais intensa que o everolimo. Observou-se ainda que o temsirolimus pode levar à apoptose tardia ou necrose após 24h de tratamento na linhagem HeLa. Dessa forma, podemos inferir que os inibidores do mTOR são ativos no tratamento do câncer do colo do útero e em especial associados à radioterapia.The PI3K/AKT/mTOR signaling pathway is frequently activated in HPV-positive, squamous cell cancers, such as cervical cancer. This study aims to investigate the biological effects of mTOR inhibitors associated or not with radiotherapy in a HPV-positive cervical cancer cell (Hela). A human keratinocytes cells (HaCaT) was used as control. Temsirolimus, everolimus, resveratrol, curcumin and EGCG were the mTOR inhibitors evaluated. The MTT cell viability assay was performed in order to obtain the dose response curves and the IC50 for each mTOR inhibitor. As comparison among the mTOR inhibitors, a cell viability analysis was done in a fixed concentration of 50µM for the diet derived mTOR inhibitors and 10µM for everolimus and temsirolimus. The cells were also pre-treated with the mTOR inhibitors at IC50 followed by radiotherapy to evaluate the radiosensitization. The scratchy assay was performed with curcumin and everolimus. The cell death profile after treatment of HeLa cells with temsirolimus, resveratrol and curcumin was assessed with flow cytometry. The results showed that the rapanalogs in a a fixed concentration of 10 µM and the diet derived mTOR inhibitors in a fixed concentration of 50µM caused mild and moderate citotoxicity in both cell lines. The association of mTOR inhibitors with radiotherapy resulted in more intense citotoxicity than radiotherapy alone. Curcumin interferes more with wound closure than everolimus in both cell lines. After 24h, temsirolimus treatment can cause late apoptosis or necrosis in HeLa cells. Based on these data, mTOR inhibitors are active in thre treatment of cervical cancer, specially in association with radiotherapy

Primary Peritoneal Carcinosarcoma in a Breast Cancer Patient Harboring a Germline BRCA2 Pathogenic Variant: Case Report

Malignant mixed müllerian tumor (MMMT) is a rare neoplasm, consisting of carcinomatous (epithelial) and sarcomatous (mesenchymal) components that most commonly arise in the endometrium and more infrequently in the ovaries, fallopian tube, cervix, and vagina. Primary peritoneal carcinosarcoma (PPCS) is an extremely rare extragenital presentation of MMMT. Although the occurrence of breast cancer and epithelial ovarian carcinoma in association with BRCA pathogenic variants is firmly established, the etiologic role of these genes in the development of other tumor types is less well known. Here, we present a rare case of PPCS in a 42-year-old Brazilian woman with a BRCA2 pathogenic variant, c.2808_2811del (NM_000059.3). The patient developed metastatic breast cancer at the age of 37 and underwent a risk-reducing bilateral salpingo-oophorectomy 2 years later. She was then diagnosed with PPCS 3 years after the risk-reducing surgery. She underwent treatment with surgery, chemotherapy, and targeted therapy but passed away almost 5 years after the second primary tumor diagnosis. To our knowledge, this is the first case of peritoneal carcinosarcoma described in a BRCA2 pathogenic variant carrier, and its report leads to a better understanding of the disease’s molecular features and possible therapeutic approaches

In vivo and in vitro effects of curcumin on head and neck carcinoma: a systematic review

Head and neck squamous cell carcinoma (HNSCC) contributes globally to a great number of deaths and morbidity, in spite of new therapeutic strategies. There is a great need of new drugs that are significantly effective and less deleterious to the patients' general health. In this sense, phytotherapy is a tendency, with results pointing to its use as a chemo-preventive and adjuvant therapy. Therefore, the objective of this systematic review was to investigate the effects of curcumin on proliferation and survival of HNSCC. The search was conducted on six databases: Cochrane, LILACS, EMBASE, MEDLINE, PubMed, and Web of Science. In vitro and in vivo studies that evaluated the effects of curcumin on cell viability, tumor growth, cell cycle and/or cell death pattern in HNSCC cell lines or animal models were selected. Of the 525 initially gathered studies, 30 met the inclusion criteria. These studies demonstrated that curcumin induces cytotoxicity, apoptosis (via intrinsic pathway), and cell cycle arrest in G2/M phase in HSNCC cell lines. It also reduces tumor measurements in animal models. These events were mostly studied through MTT assay, flow cytometry, and cell cycle- and apoptosis-related proteins expression. This systematic review demonstrated that curcumin is effective on HNSCC cell proliferation and survival, reinforcing the currently available evidence that curcumin could be an adjuvant drug in HNSCC treatment.46132

Using an Untargeted Metabolomics Approach to Identify Salivary Metabolites in Women with Breast Cancer

Metabolic alterations are a hallmark of the malignant transformation in cancer cells, which is characterized by multiple changes in metabolic pathways that are linked to macromolecule synthesis. This study aimed to explore whether salivary metabolites could help discriminate between breast cancer patients and healthy controls. Saliva samples from 23 breast cancer patients and 35 healthy controls were subjected to untargeted metabolomics using liquid chromatography-quadrupole time-of-flight mass spectrometry and a bioinformatics tool (XCMS Online), which revealed 534 compounds, characterized by their retention time in reverse-phase liquid chromatography and by the m/z ratio detected, that were shared by the two groups. Using the METLIN database, 31 compounds that were upregulated in the breast cancer group (p < 0.05) were identified, including seven oligopeptides and six glycerophospholipids (PG14:2, PA32:1, PS28:0, PS40:6, PI31:1, and PI38:7). In addition, pre-treatment and post-treatment saliva samples were analyzed for 10 patients who experienced at least a partial response to their treatment. In these patients, three peptides and PG14:2 were upregulated before but not after treatment. The area under the curve, sensitivity, and specificity for PG14:2 was 0.7329, 65.22%, and 77.14%, respectively. These results provide new information regarding the salivary metabolite profiles of breast cancer patients, which may be useful biomarkers

Germline molecular data in hereditary breast cancer in Brazil: Lessons from a large single-center analysis.

Brazil is the largest country in South America and the most genetically heterogeneous. The aim of the present study was to determine the prevalence of germline pathogenic variants (PVs) in Brazilian patients with breast cancer (BC) who underwent genetic counseling and genetic testing at a tertiary Oncology Center. We performed a retrospective analysis of the medical records of Brazilian patients with BC referred to genetic counseling and genetic testing between August 2017 and August 2019. A total of 224 unrelated patients were included in this study. Premenopausal women represented 68.7% of the cohort. The median age at BC diagnosis was 45 years. Multigene panel testing was performed in 219 patients, five patients performed single gene analysis or family variant testing. Forty-eight germline PVs distributed among 13 genes were detected in 20.5% of the patients (46/224). Eighty-five percent of the patients (91/224) fulfilled NCCN hereditary BC testing criteria. Among these patients, 23.5% harbored PVs (45/191). In the group of patients that did not meet NCCN criteria, PV detection rate was 3% (1/33). A total of 61% of the patients (28/46) harbored a PV in a high-penetrance BC gene: 19 (8.5%) BRCA1/2, 8 (3.5%) TP53, 1 (0.5%) PALB2. Moderate penetrance genes (ATM, CHEK2) represented 15.2% (7/46) of the positive results. PVs detection was statistically associated (p<0.05) with BC diagnosis before age 45, high-grade tumors, bilateral BC, history of multiple primary cancers, and family history of pancreatic cancer. According to the current hereditary cancer guidelines, 17.4% (39/224) of the patients had actionable variants. Nine percent of the patients (20/224) had actionable variants in non-BRCA genes, it represented 43.5% of the positive results and 51.2% of the actionable variants. Considering the observed prevalence of PVs in actionable genes beyond BRCA1/2 (9%, 20/224), multigene panel testing may offer an effective first-tier diagnostic approach in this population

PARP inhibitors as first-line maintenance therapy in ovarian cancer: recommendations from an expert panel from Brazil

To report consensus recommendations on the current role of poly(ADP-ribose) polymerase (PARP) inhibitors in the front-line management of patients with epithelial ovarian cancer (EOC) in the healthcare setting of Brazil. The expert panel convened in March 2021 and comprised 20 medical oncologists focus on gynecological oncology. The panel answered anonymously and based on scientific evidence a total of 67 questions. The panel reached consensus (at least 75% of votes for the same recommendation) or majority vote (50% to 74.9%) for the majority of questions that addressed: (1) who and when to test for BRCA mutations or homologous recombination deficiency (2) what test should be used; (3) when should maintenance PARP inhibitor therapy be indicated; (4) which PARP inhibitor should be used; (5) when should bevacizumab be combined; and (6) toxicity management. The current recommendations may help Brazilian practitioners to improve the use of PARP inhibitors in front-line management of EOC

Cost-effectiveness analysis of Oncotype DX from a Brazilian private medicine perspective: a GBECAM multicenter retrospective study

Background: Oncotype DX (ODX) is a validated assay for the prediction of risk of recurrence and benefit of chemotherapy (CT) in both node negative (N0) and 1–3 positive nodes (N1), hormone receptor positive (HR+), human epidermal growth factor receptor 2-negative (HER2−) early breast cancer (eBC). Due to limited access to genomic assays in Brazil, treatment decisions remain largely driven by traditional clinicopathologic risk factors. ODX has been reported to be cost-effective in different health system, but limited data are available considering the reality of middle-income countries such as Brazil. We aim to evaluate the cost-effectiveness of ODX across strata of clinical risk groups using data from a dataset of patients from Brazilian institutions. Methods: Clinicopathologic and ODX information were analyzed for patients with T1–T3, N0–N1, HR+/HER2− eBC who had an ODX performed between 2005 and 2020. Projections of CT indication by clinicopathologic criteria were based on binary clinical risk categorization based on the Adjuvant! Algorithm. The ODX score was correlated with the indication of CT according to TAILORx and RxPONDER data. Two decision-tree models were developed. In the first model, low and high clinical risk patients were included while in the second, only high clinical risk patients were included. The cost for ODX and CT was based on the Brazilian private medicine perspective. Results: In all, 645 patients were analyzed; 411 patients (63.7%) had low clinical risk and 234 patients (36.3%) had high clinical risk disease. The ODX indicated low (25) risk in 119 (18.4%), 415 (64.3%), and 111 (17.2%) patients, respectively. Among 645 patients analyzed in the first model, ODX was effective (5.6% reduction in CT indication) though with an incremental cost of United States Dollar (US$) 2288.87 per patient. Among 234 patients analyzed in the second model (high clinical risk only), ODX led to a 57.7$ 4350.66 per patient. Conclusions: Our study suggests that ODX is cost-saving for patients with high clinical risk HR+/HER2− eBC and cost-attractive for the overall population in the Brazilian private medicine perspective. Its incorporation into routine practice should be strongly considered by healthcare providers

Key issues in the management of cervical cancer: consensus recommendations by a Brazilian expert panel

Objective: We report the results of a panel of Brazilian experts and provide recommendations for the management of these patients. Material and Methods: The panel convened composed by 28 local opinion leaders, addressed 59 multiple-choice questions taking into account the published scientific literature and their own clinical experience. The level of agreement among panel members was qualified as (1) consensus, when at least 75% of the voting panel members; (2) majority vote (50%-74.9%); or (3) less than majority vote. Results: There was at least majority vote for eight of 10 questions on staging and follow-up; for 14 of 23 questions on the treatment of early-stage disease; for 12 of 14 questions related to the treatment of locally-advanced disease; and for seven of the 12 questions related to the treatment of recurrent/metastatic disease. Conclusion: The current recommendations may help practitioners from Brazil and other countries to improve the care they provide to patients

O Protagonismo Infantojuvenil nos Processos Educomunicativos

Neste volume “O protagonismo infantojuvenil nos processos educomunicativos”, reunimos 53 artigos que transitam sobre a temática do protagonismo infantojuvenil em diversas experiências e processos educomunicativos e para facilitar sua leitura e busca por temas de seu interesse, eles estão organizados em 8 capítulos que abordam a educomunicação a partir do fazer das crianças e da apropriação da produção midiática.&nbsp;Expressão artística, rádio, vídeo, jornalismo, cultura digital, redes sociais entre outros são os temas abordados pelos autores destes trabalhos. convidamos o leitor&nbsp;a mergulhar nesta jornada educomunicativa, vivendo e revivendo junto conosco essas experiências vividas por outros, refletindo em cada texto sobre como estamos, como evoluímos e como seguimos os passos daqueles que com sua ousadia, amor e luta elaboraram os fundamentos da educomunicação