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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.
Resumo Objetivo O presente estudo avaliou o perfil de mutações germinativas presentes em pacientes submetidas a aconselhamento genético para avaliação de risco para câncer de mama (CM), câncer de ovário (OC) e câncer de endométrio (CE) com possível padrão hereditário. Métodos Foram analisados os prontuários de 382 pacientes que realizaram aconselhamento genético após consentimento informado. Um total de 55,76% dos pacientes (213/382) eram sintomáticos (história pessoal de câncer), e 44,24% (169/382) eram assintomáticos (ausência da doença). As variáveis analisadas foram idade, sexo, naturalidade, história pessoal ou familiar de CM, OC, CE bem como outros tipos de câncer associados a síndromes hereditárias. As diretrizes de nomenclatura da Human Genome Variation Society (HGVS) foram usadas para nomear as variantes e seu significado biológico foi determinado pela comparação de 11 bancos de dados. Resultados Identificamos 53 mutações distintas: 29 variantes patogênicas, 13 variantes de significado indeterminado e 11 benignas. As mutações mais frequentes foram BRCA1 c.470_471delCT, BRCA1 c.4675 + 1G > T e BRCA2 c.2T > G. Além disso, 21 variantes parecem ter sido descritas pela primeira vez no Brasil. Além das mutações BRCA1/2, foram encontradas variantes em outros genes relacionados a síndromes hereditárias que predispõem a cânceres ginecológicos. Conclusão Este estudo permitiu conhecer melhor as principais mutações identificadas nas famílias do estado de Minas Gerais e demonstra a necessidade de avaliar a história familiar de câncer não ginecológico para avaliação do risco de CM, OC e CE. Além disso, é um esforço que contribui com estudos populacionais para avaliar o perfil de mutações de risco para câncer no Brasil.
Subject(s)
Humans , Female , Breast Neoplasms/prevention & control , Risk Factors , Endometrial Neoplasms/prevention & control , Genetic Counseling , Genital Neoplasms, Female/prevention & control , Genetic Diseases, InbornABSTRACT
ABSTRACT Objective To investigate the expression of human papillomavirus (HPV), p16, p53, and p63 in non-schistosomiasis-related squamous cell carcinoma of the bladder and to develop an accurate and automated tool to predict histological classification based on clinicopathological features. Methods Twenty-eight patients with primary bladder pure squamous cell carcinoma who underwent cystectomy or transurethral resection of bladder tumor (TURBT) for bladder cancer between January 2011 and July 2017 were evaluated. Clinical data and follow-up information were obtained from medical records. Formalin-fixed, paraffin-embedded surgical specimens were used for immunohistochemical staining for p16, p53, and p63. Human papillomavirus detection was evaluated by PCR. Statistical analysis was performed, and statistical significance was set at p<0.05. Finally, decision trees were built to classify patients' prognostic features. Leave-one-out cross-validation was used to test the generalizability of the model. Results Neither direct HPV detection nor its indirect marker (p16 protein) was identified in most cases. The absence of p16 was correlated with less aggressive histological grading (p=0.040). The positive p16 staining detection found only in pT1 and pT2 cases in our sample suggests a possible role for this tumor suppressor protein in the initial stages of bladder squamous cell carcinoma. The decision trees constructed described the relationship between clinical features, such as hematuria/dysuria, the level of tumor invasion, HPV status, lymphovascular invasion, gender, age, compromised lymph nodes, and tumor degree differentiation, with high classification accuracy. Conclusion The algorithm classifier approach established decision pathways for semi-automatic tumor histological classification, laying the foundation for tailored semi-automated decision support systems for pathologists.
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OBJECTIVES: 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. METHODS: 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). RESULTS: 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. CONCLUSION: 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.
Subject(s)
Humans , Female , Ovarian Neoplasms , Cisplatin , Apoptosis , Cell Line, Tumor , Antineoplastic AgentsABSTRACT
Dentre as patologias do trato genital feminino, o câncer epitelial de ovário (CEO) representa a sétima causa mais relevante de morbidade e mortalidade no mundo. Apesar dos avanços no tratamento dessa neoplasia, não houve melhorias equivalentes nas tax as de sobrevida das mulheres afetadas. O presente estudo visa analisar o perfil de expressão e metilação dos genes receptor es tipo TRAIL ( TRAIL-R2 e TRAIL-R3) , CASPASE- 8 e BCL -2 em pacientes com CEO a fim de prover uma melhor compreensão da biologia molecular deste tumor e da predição de respostas terapêuticas numa medicina person alizada. Para as análises, foram coletadas amostras de pacientes atendidas no Hospital Vera Cruz, em Belo Horizonte, Minas Gerais. Estas amostras foram destinadas à extração simultânea do R NA e DNA genômico pelo reagente TR Izol®. A expressão gênica de TRAIL-R2, TRAIL-R3, BCL-2 e CASPASE-8 foi avaliada por meio da reação em cadeia da polimerase (PCR) quantitativa em tempo real e o padrão de metilação dos promotores de TRAIL-R2, TRAIL-R3 e C ASPASE-8 foi analisado por PCR metilação específica. Observou-se que o grupo de pacientes com CEO metastático superexpressou todos os genes avaliados, enquanto que, pacientes com cistoadenoma seroso ovariano apenas não superex pressar am BCL-2. O grupo de pacientes com CEO primário expressou níveis reduzidos dos marcadores antiapoptóticos, TRAIL-R3 eBCL-2¸ que aliados a superex pressão de CASPASE -8, sugerem completamente não metiladas para o marcador TRAIL-R3 e que difer enças significativas foram encontradas entre a metilação presente nas ilhas CpG dos promotores de TRAILR3 eCASPASE-8 e os grupos histológicos. Entretanto, esse evento não se correlacionou com o silenciamento epigenético desses genes, o que sugere o envolvimento de outros mecanismos de controle transcricional...
The epithelial ov arian cancer (EOC) represents the seventh most important cause of morbidity and mortality worldwide among the female genital tract diseases. Despite the advances in the treatment of this malignancy, there were no equivalent improvements in the survival rates of affected women. The present study aims to analyze the expression and methylation profiles of TRAIL receptors ( TRAIL-R2 and TRAIL-R3 ), CASPASE-8 and BCL-2 genes in p atients with EOC in order to provide a better understanding of the molecular biology of this pathology and to predict therapeutic responses in a personalized medicine. Samples were collected from patients treated at Vera Cruz Hospital in Belo Horizonte, state of Minas Gerais. Those samples were used for simultaneous extraction of RNA and genomic DNA b y TRIzol ® reagent. The genetic expression of TRAIL-R2, TRAIL-R3, BCL-2 and CASPASE-8 were evaluated by realtime quantitative polymerase chain reaction and the meth ylation profile of TRAIL-R2, TRAIL-R3 and CASPASE-8 promoters were analyzed b y methylation specific PCR. Higher expression levels of all four markers were observed in metastatic EOC. In contrast, serous cystoad enoma overexpressed all markers, ex cept for BCL-2. The primary tumors underexpressed anti-apoptotic gen es, TRAIL-R3 and BCL-2 , and overexpressed proapoptotic genes C ASPASE -8 which suggest a genetic signature in TRAIL signaling pathway in EOC...