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Biomédica (Bogotá) ; 41(4): 773-786, oct.-dic. 2021. tab, graf
Article in English | LILACS | ID: biblio-1355749


Abstract | Introduction: Next Generation Sequencing (NGS) is cost-effective and a faster method to study genes, but its protocol is challenging. Objective: To analyze different adjustments to the protocol for screening the BRCA genes using Ion Torrent PGM sequencing and correlate the results with the number of false positive (FP) variants. Materials and methods: We conducted a library preparation process and analyzed the number of FP InDels, the library concentration, the number of cycles in the target amplification step, the purity of the nucleic acid, the input, and the number of samples/Ion 314 chips in association with the results obtained by NGS. Results: We carried out 51 reactions and nine adjustments of protocols and observed eight FP InDels in homopolymer regions. No FP Single-Nucleotide Polymorphism variant was observed; 67.5% of protocol variables were jointly associated with the quality of the results obtained (p<0.05). The number of FP InDels decreased when the quality of results increased. Conclusion: The Ion AmpliSeq BRCA1/BRCA2 Community Panel had a better performance using four samples per Ion-314 chip instead of eight and the optimum number of cycles in the amplification step, even when using high-quality DNA, was 23. We observed better results with the manual equalization process and not using the Ion Library Equalizer kit. These adjustments provided a higher coverage of the variants and fewer artifacts (6.7-fold). Laboratories must perform internal validation because FP InDel variants can vary according to the quality of results while the NGS assay should be validated with Sanger.

Resumen | Introducción. La secuenciación de nueva generación es un método rentable y rápido para el estudio de los genes, pero su protocolo entraña desafíos. Objetivo. Investigar diferentes ajustes del protocolo de selección de los genes BRCAmediante secuenciación de Ion Torrent PGM™ y correlacionar los resultados con el número de variantes de falso positivo. Materiales y métodos. El proceso de preparación de la biblioteca, el número de falsos positivos InDels, la concentración de la biblioteca, el número de ciclos en el paso de amplificación de objetivos, la pureza del ácido nucleico, la entrada y el número de muestras por chip del Ion-314 se analizaron en asociación con los resultados obtenidos por secuenciación de nueva generación secuenciación de nueva generación. Resultados. Se hicieron 51 reacciones y nueve ajustes de los protocolos, y se observaron ocho falsos positivos InDels en las regiones de homopolímeros. No se observó ninguna variante de polimorfismo de nucleótido simple falso positivo. En 67,5 % de los casos, las variables de protocolo en su conjunto se asociaron con la calidad de los resultados obtenidos (p<0,05). El número de falsos positivos InDels disminuyó al aumentar la calidad de los resultados. Conclusiones. El panel comunitario Ion AmpliSeq BRCA1/BRCA2 tuvo un mejor rendimiento, con cuatro muestras por chip Ion-314 en lugar de ocho, y el número de ciclos en el paso de amplificación, incluso con ADN de alta calidad, fue mejor con 23. Se observaron mejores resultados con el proceso de ecualización manual y sin el uso del kit Ion Library Equalizer. Estos ajustes proporcionaron una mayor cobertura de las variantes y menos artefactos. Los laboratorios deben realizar la validación interna porque las variantes de falsos positivos InDel pueden variar según la calidad de los resultados. La secuenciación de próxima generación debe validarse con Sanger.

DNA , High-Throughput Nucleotide Sequencing , Sequence Analysis , Genes, BRCA1 , Genes, BRCA2
Acta sci. vet. (Impr.) ; 49: Pub. 1827, 2021. tab
Article in Portuguese | LILACS, VETINDEX | ID: biblio-1363756


Mammary tumors (MTs) in bitches are similar to breast cancers in women. Thus, they can be used as a model for human breast cancer and findings can be extrapolated for use in human medicine. BRCA1 is a tumor suppressor gene. When the gene has a mutation, it cannot repair damaged DNA, which causes genetic instability and tumorigenesis. Therefore, we aimed to study the frequency of single nucleotide polymorphisms (SNPs) in the BRCA1 gene that are associated with distinct histological types of malignant MT in bitches. The study population consisted of 91 bitches, including a control group of 6 animals with healthy mammary glands and 85 animals with MTs. All animals underwent a presurgery evaluation consisting of a questionnaire administered to the person responsible for the animal, a physical examination, collection of peripheral blood for hematological and serum biochemistry evaluations, an electrocardiogram, and a preanesthesia evaluation. In addition, distant metastasis was studied via chest radiography and abdominal ultrasound. After evaluations were complete, the animals that could undergo surgery were administered general anesthesia and underwent a mastectomy or mammary gland sample collection. Histopathological examination and molecular analysis were performed to identify mutations in the BRCA1 gene. Histopathological examinations found 10 different types of malignant tumors in 36 sick animals. Tumor samples plus samples from the 6 control animals were subjected to DNA extraction, polymerase chain reaction (PCR) analysis, and genetic sequencing. The tumor with the highest incidence (33.33%) was a complex carcinoma, followed by carcinoma in mixed tumor (13.88), tubular carcinoma (13.88) and carcinosarcoma (13.88). Molecular analysis revealed 3 different SNP points in 5 samples (4006G>A, 3619A>G, and 3761C>T). The allelic variant 4006G>A (1/36) resulted in the alteration of the amino acid valine by isoleucine (V1336 I). The mutation 3619A>G (2/36) inserted the amino acid alanine instead of threonine (T1207 A). The mutation 3761C>T (2/36) led to the alteration of the amino acid serine by phenylalanine (S1254 F), a mutation for which there are no published reports. The histological types that showed BRCA1 mutations were complex carcinoma (1/5), carcinoma in mixed tumor (1/5), papillary carcinoma (1/5) and tubular carcinoma (2/5). Software analysis identified the new SNP (nucleotide 3761) in BRCA1 and 2 point mutations in nucleotides 4006 and 3619 and responsible for genetic instability. The development of breast cancer is caused by many endogenous and exogenous factors. The results of our study show that these factors have a greater presence in female, mixed breed, uncastrated, and older dogs, confirming the data in the veterinary literature. In the present study, we found different histological types of malignant breast tumors with mutations in the BRCA1 gene, as other authors have reported. However, we also found the mutation 3761C>T, which, to the best of our knowledge, has not been reported in the literature. This shows the need for studies in veterinary medicine that assess mutations in the BRCA1 gene and the most common histological types. In conclusion, SNPs in the BRCA1 gene cause genetic instability, resulting in additional mutations that lead to the development of breast tumors. They are point mutations that affect transcription, resulting in truncated proteins. These proteins may have a loss of function, leading to carcinogenesis.(AU)

Animals , Female , Dogs , Mammary Neoplasms, Animal/genetics , Mammary Neoplasms, Animal/diagnostic imaging , Genes, BRCA1 , Polymorphism, Single Nucleotide/genetics , Dog Diseases/genetics , Dogs
Biomédica (Bogotá) ; 40(1): 185-194, ene.-mar. 2020. tab, graf
Article in Spanish | LILACS | ID: biblio-1089114


Introducción. El cáncer de mama es un problema mundial de salud pública; entre el 5 y el 10 % de los casos presentan agregación familiar, lo que se explicaría por la presencia de mutaciones en genes de alto riesgo como el BRCA1 y el BRCA2. El origen fundador de la deleción BRCA1 3450del4 en Colombia ya fue reportado. Objetivo. Hacer un análisis descriptivo de seis familias del del Tolima y del Huila con la deleción BRCA1 3450del4 de la asociación de la mutación germinal, con el cáncer de mama y la agregación familiar. Materiales y métodos. Se hizo un estudio descriptivo y transversal de seis casos índice con cáncer de mama positivos para BRCA1 3450del4, que cumplían tres de los criterios establecidos por Jalkh, et al. A partir de la información de las entrevistas, se realizaron los árboles genealógicos (GenoPro™, versión 2016). Se tipificó la mutación en familiares sanos y afectados que aceptaron participar. Resultados. De los 78 individuos seleccionados por conveniencia en las seis familias, 30 presentaron la mutación BRCA1 3450del4; de ellos, seis tenían cáncer de mama, uno, cáncer de ovario, uno, cáncer de mama y ovario, y otro, cáncer de próstata; 21 no presentaban neoplasias. De los 30 individuos portadores de la variante patogénica, seis eran hombres y 24 mujeres, 13 de ellas menores de 30 años. Conclusiones. En este estudio se confirmó la asociación de la deleción BRCA1 3450del4 con el cáncer de mama de agregación familiar.

Introduction: Breast cancer is a worldwide public health problem; between 5% and 10% of the cases present familial aggregation explained by genes of high risk such as BRCA1and BRCA2. The founding origin of the deletion BRCA1 3450del4 in Colombia has been previously reported. Objective: To carry out in six families from Tolima and Huila departments a descriptive analysis of the presence of the BRCA1 3450del4 mutation associated with breast cancer and familial aggregation. Materials and methods: We conducted a descriptive and cross-sectional study of six index cases with breast cancer positive for BRCA1 3450del4 that fulfilled three of the criteria established by Jalkh, et al. The genealogical trees were made using the information of the interview data (GenoPro™, version 2016). The mutation was typified in healthy and affected relatives who agreed to participate. Results: Thirty of the 78 individuals selected by convenience in the six families presented the mutation BRCA1 3450del4 six of whom developed breast cancer, one, ovarian cancer, one ovarian and breast cancer, and one prostate cancer; 21 did not present any type of neoplasm at the time of the study. Of the 30 individuals carrying the pathogenic variant, six were men, 24 were women, and 13 of these were under 30. Conclusions: In this study of families with the deletion BRCA1 3450del4 in Tolima and Huila we confirmed its association with familial aggregation of breast cancer.

Breast Neoplasms/genetics , Chromosome Deletion , Genes, BRCA1 , Mutation
Article in English | WPRIM | ID: wpr-763185


PURPOSE: We investigated the clinical relevance and spectrum of BRCA1/2 mutations in Korean ovarian cancer (KoOC) patients. MATERIALS AND METHODS: Two hundred seventy-nine KoOC patients were enrolled from three university hospitals between 2012 and 2017. Their peripheral blood samples were obtained for BRCA1/2 mutation analysis by direct sequencing. Clinicopathological characteristics were retrospectively reviewed, and spectrum analyses of BRCA1/2 mutation were assessed by systematic literature review. RESULTS: Frequency of BRCA1/2 mutations was 16.5% in KoOC patients. BRCA1/2 mutations were significantly associated with family history of breast/ovarian cancer (pT of BRCA2 in KoBC). CONCLUSION: The clinical relevance of BRCA1/2 mutations in KoOC patients was confirmed but that of early age-of-onset was not. Possible inconsistency in the ratio of BRCA1-to-BRCA2 mutations and the most common mutation between KoOC and KoBC may probably suggest presence of mutation sequence-associated penetrance tendency in hereditary Korean breast and ovarian cancer. These data may provide insights for optimal genetic counseling and prophylactic treatment for at-risk relatives of KoOC patients.

Asians , Breast , Breast Neoplasms , Genes, BRCA1 , Genes, BRCA2 , Genetic Counseling , Germ-Line Mutation , Gynecology , Hospitals, University , Humans , Obstetrics , Ovarian Neoplasms , Penetrance , Retrospective Studies
Article in English | WPRIM | ID: wpr-760678


OBJECTIVE: BRCA mutational status is important in the management of ovarian cancer, but there is a lack of evidence supporting genetic testing in Asian populations. This study was performed to investigate the prevalence and prognostic outcomes of BRCA1/2 mutation and variant of unknown significance (VUS) in Korean patients diagnosed with epithelial ovarian cancer (EOC). METHODS: Among patients newly diagnosed with EOC between January 2007 and January 2017, those tested for germline BRCA1/2 mutation were studied, regardless of family history. Overall survival (OS) and progression-free survival (PFS) were compared between the patients with and without BRCA1/2 mutation and VUS. RESULTS: A total of 313 patients underwent BRCA testing: 88 patients had a BRCA1/2 mutation and 48 patients had a BRCA1/2 VUS (28.1% and 15.3%, respectively). There were no significant associations between BRCA1/2 mutation, BRCA1/2 wild-type, or BRCA1/2 VUS with age at diagnosis, histologic distribution, or residual disease status after primary cytoreductive surgery. BRCA1 mutation, including BRCA1 VUS, showed no difference in PFS or OS compared to BRCA1 wild-type. In contrast, BRCA2 mutation showed longer PFS than that of BRCA2 wild-type (P=0.04) or BRCA2 VUS (P=0.02). BRCA2 mutation, including BRCA2 VUS, did not show any difference in OS compared to BRCA2 wild-type. CONCLUSION: BRCA mutation and BRCA VUS had similar clinical characteristics and survival outcomes, except that BRCA2 mutation showed better PFS. The results of this study will help to understand the prognostic significance of BRCA mutation and VUS in Korean patients.

Asians , Diagnosis , Disease-Free Survival , Genes, BRCA1 , Genes, BRCA2 , Genetic Testing , Humans , Korea , Ovarian Neoplasms , Prevalence
São Paulo; s.n; 2019. 140 p. ilust, tabelas, quadros.
Thesis in Portuguese | LILACS, Inca | ID: biblio-1179202


O Câncer de Mama Triplo-Negativo (CMTN), caracterizado pela perda de expressão dos receptores de estrógeno (RE), de progesterona (RP) e pela não super-expressão/amplificação do receptor do fator de crescimento epidermal humano do tipo 2 (HER-2), é considerado um subtipo bastante agressivo e heterogêneo molecularmente. Clinicamente, o CMTN apresenta prognóstico ruim, altas taxas de recorrência e menor sobrevida global em relação aos outros subtipos de câncer de mama. Ele corresponde a aproximadamente 15% dos casos de câncer de mama e não apresenta nenhuma terapia alvo efetiva. Mutação patogênica germinativa nos genes BRCA1 e BRCA2 leva a um aumento de risco para o desenvolvimento de Câncer de Mama e Ovário, sendo que mutação germinativa em BRCA1 está associada ao desenvolvimento de CMTN, especialmente em pacientes diagnosticadas antes dos 50 anos. A deficiência de BRCA1 leva ao mecanismo ineficiente do reparo do DNA e ao desenvolvimento do tumor. Recentemente, nosso grupo classificou as pacientes diagnosticadas com CMTN em grupo Hereditário (com mutação patogênica germinativa em BRCA1) e grupo Esporádico (sem mutação germinativa em BRCA1). Estes foram ainda classificados em tumores BRCA1-deficiente (com hipermetilação no promotor de BRCA1) e em tumores BRCA1-proficiente (sem hipermetilação no promotor de BRCA1 e sem mutação germinativa em BRCA1). As diferenças moleculares entre esses grupos de CMTN são de grande interesse clínico e biológico, embora ainda não sejam conhecidas. Dentro desse contexto, nosso objetivo foi investigar o perfil transcricional de CMTN sob diferentes aspectos de deficiência de BRCA1. Para isso, foi avaliado o sequenciamento do RNA (RNA-Seq) de 37 casos de CMTN em idade jovem (≤ 50 anos), sendo 9 casos Hereditário (com mutação patogênica germinativa em BRCA1) e 28 casos Esporádico. Dos casos Esporádicos, 9 foram BRCA1-deficiente (com o promotor de BRCA1 hipermetilado) e 19 BRCA1-proficiente (com o promotor de BRCA1 não hipermetilado). Utilizamos o RNA total a partir das 37 amostras tumorais e 25 amostras normais pareadas adjacentes ao tumor. As bibliotecas de cDNA foram construídas a partir do RNA total através do método de depleção do rRNA e sequenciadas na plataforma NextSeq (Illumina). Para a normalização dos dados de expressão de cada gene, foi aplicada a medida FPKM. Os critérios para a obtenção dos genes diferencialmente expressos (GDEs), nas amostras tumorais em relação às normais, foram fold change ≥ 4,0 e ≤ - 4,0 e p valor ajustado ≤ 0,05). Para classificação molecular dos tumores TN, foi utilizada a ferramenta online TNBCtype. As curvas de sobrevida global de acordo com essa classificação molecular foram calculadas através do método de Kaplan-Meier. Para verificar os processos biológicos envolvidos com a tumorigênese do CMTN, os GDEs foram submetidos a uma análise funcional in silico através do programa Ingenuity Pathway Analysis (IPA). Em média, 48.627.204 milhões de sequências foram geradas por amostra, das quais 79,5% foram mapeadas no genoma humano referência, revelando, em média, 15.071 genes expressos com pelo menos 10 sequências única mapeada por amostra. O perfil transcricional das amostras CMTN permitiu a classificação nos 7 subtipos moleculares de CMTN, sendo a maioria das amostras classificadas no subtipo imunomodulador (IM) e mesenquimal (M). No grupo das amostras BRCA1-deficiente, observamos um predomínio do subtipo IM enquanto que, nas amostras BRCA1-proficiente, houve um maior número de amostras classificadas em M. Não observamos diferença nas curvas de sobrevida global entre os dois grupos de amostras. Porém, os resultados mostraram que o subtipo IM parece ter uma tendência de melhor sobrevida global comparado com os outros subtipos, independente do status de BRCA1. A expressão diferencial das amostras tumorais em relação às amostras normais no grupo de CMTN Hereditário revelou 1965 GDEs, sendo 589 mais expressos e 1376 menos expressos no tumor; e, no grupo de CMTN Esporádico, a análise revelou 1837 GDEs, sendo 645 mais expressos e 1192 menos expressos no tumor. A partir dos GDEs de cada grupo, a análise do IPA mostrou que os dois grupos apresentaram vias canônicas enriquecidas comuns significantemente ativadas e envolvidas, de forma geral, com a Regulação do ciclo celular. Entretanto, as vias canônicas significantemente inibidas mostraram-se exclusivas em cada grupo: a via de Sinalização do receptor do glutamato foi a mais significativa (z-score = -2,12) nas amostras de CMTN Hereditário; e a via de Ativação de LXR/RXR foi a mais significativa (z-score = -2,98) nas amostras CMTN Esporádico. Além disso, observamos reguladores transcricionais significantemente ativados e inibidos exclusivamente em cada grupo. Os genes TNF e E2F1 foram os mais significantemente ativados apenas nas amostras de CMTN Hereditário (z-score = 4,06) e de CMTN Esporádico (z-score = 2,22), respectivamente. Por outro lado, o microRNA mir-21 e o gene PPARG foram os reguladores inibidos exclusivamente nas amostras CMTN Hereditário (-score = -4,68) e nas amostras CMTN Esporádico (z-score = -4,24), respectivamente. Esse estudo, de forma geral, revelou potenciais vias e genes reguladores de cascatas de sinalização envolvidos na tumorigênese do CMTN no contexto da deficiência de BRCA1, contribuindo na elucidação da complexidade funcional de tumores TN

Triple-negative breast cancer (TNBC), characterized by lack of expression of the estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2), results in aggressive biology, early peak of recurrence, and shorter overall survival than other subtypes. It comprises approximately 15% of breast cancer cases and yet there is no effective therapy. Mutations in the BRCA1 and BRCA2 genes are associated with increased risk of breast and ovarian cancers since germline mutation in BRCA1 is associated with the development of TNBC, especially in patients diagnosed before age 50. BRCA deficiency leads to impaired DNA repair and tumor development. Recently, our group classified TNBC patients into hereditary BRCA1-mutated and sporadic BRCA1-proficient. These were further classified into BRCA1-deficient tumors (with BRCA1 promotor hypermethylation) and BRCA1-proficient tumors (no BRCA1 promoter hypermethylation neither BRCA1 germline mutation). Molecular differences between hereditary and sporadic TNBC groups are clinically and biologically interesting although it remains unclear. In this context, we aimed to investigate the transcriptional profile of TNBC-associated or not with BRCA1 deficiency. For that, RNA sequencing (RNA-seq) from 37 early-onset TNBC (≤ 50 years old) was evaluated, comprising 9 Hereditary (with BRCA1 germline pathogenic mutation) and 28 Sporadic cases, of which 9 BRCA1-deficient and 19 BRCA1-proficient. Total RNA from 37 tumors samples and 25 adjacent normal samples of paired cases were used to constructed RNAseq libraries by depleting ribosomal RNA and sequenced on Illumina NextSeq 500 platform. Expression values were normalized by FPKM. Differentially expressed genes (DEGs) between tumor and normal samples were obtained using fold-change ≥ |4| and p value adjusted ≤ 0.05 as statistical criteria. The TNBC samples were subtyping using the web-based prediction tool TNBCType. Overall survival curves were calculated using the Kaplan.Meir method. IPA software was used to detect activated/inactivated canonical pathways and relevant upstream regulators, considering a z-score ≤-2.0 and ≥ 2.0, respectively. On average, 49 million reads were generated per sample, of which 79.5% were mapped to the human reference genome revealing 15,071 expressed genes with at least 10 reads per sample. From the transcriptional profile of TNBC samples we classified into seven TNBC subtypes being the majority of tumors classified as immunomodulatory (IM) and mesenchymal (M) subtype. We detected no difference in overall survival for both groups. However, trends towards better overall were observed for TNBC samples classified as IM compared with other subtypes, without associations with BRCA1 status. Differential gene expression analysis between tumor and normal samples in the hereditary group revealed 1,965 DEGs, being 589 upregulated and 1,376 downregulated; and in the sporadic group, the analysis revealed 1,837 DEGs, being 645 upregulated and 1,192 downregulated. Using the DEGs of each group, the IPA analysis revealed that Cell Cycle Regulation signaling was activated in both groups. Regarding inactivated pathways, we detected the Glutamate Receptor signaling (z-score = -2,12) in hereditary TNBC and the LXR/RXR activation in sporadic TNBC (z-score = -2,98). Also, the IPA analysis revealed relevant specific transcription regulators of each group. The TNF and E2F1 were the most significantly activated genes in hereditary- and sporadic-TNBC, respectively. On the other hand, the mir-21 and PPARG were the most significantly unique inhibited regulators in hereditary and sporadic, respectively. In general, this study unveiled potential pathways and regulatory genes for signaling cascades involved in TNBC tumorigenesis considering the deficiency of BRCA1, contributing to the elucidation of the functional complexity of the tumorigenic process of TNBC patients

Humans , Female , Adult , Middle Aged , Aged , Breast Neoplasms , Sequence Analysis, RNA , Germ-Line Mutation , Genes, BRCA1 , High-Throughput Nucleotide Sequencing , Triple Negative Breast Neoplasms/genetics
Rev. bras. anal. clin ; 50(1): 17-21, jun. 2018.
Article in Portuguese | LILACS | ID: biblio-911927


O câncer de mama é o principal câncer que atinge a população feminina no mundo, com maior taxa de incidência e mortalidade, sendo que de 5% a 10% de todos os casos são relacionados à herança de mutações genéticas. A identificação precoce dos casos de câncer de mama e ovário é importante, pois um indivíduo afetado pode herdar propriedade de antecedentes familiares que indicam uma predisposição hereditária. O efeito cancerígeno pode ocorrer quando dois genes supressores de maior importância, como BRCA1 e BRCA2, perdem suas funções nos dois alelos decorrentes de mutações na linhagem germinativa. Desta forma, foi realizada uma revisão da literatura sobre câncer de mama hereditário e suas correlações com mutações germinativas nos genes BRCA1 e BRCA2 que aumentam o risco para o desenvolvimento de câncer de mama.

Breast Neoplasms , Genes, BRCA1 , Genetic Predisposition to Disease , Genes, BRCA2
Rev. salud pública ; 20(2): 232-236, mar.-abr. 2018. tab, graf
Article in Spanish | LILACS | ID: biblio-978966


RESUMEN Objetivo Evaluar la utilidad de la cirugía reductora de riesgo en trompas y ovarios en casos con mutación del gen BRCA1 en Colombia Material y Métodos Análisis de costo-efectividad en el que se incorporan tres procesos: a. Pacientes con pruebas de tamización para cáncer de mama y ovario. b. cirugía reductora de riesgo en trompas y ovarios c. cirugía reductora de riesgo en trompas y ovarios con mastectomía bilateral. Se evalúa como desenlace la ganancia en años de supervivencia. Resultados La cohorte con cirugía reductora de riesgo en trompas y ovarios y mastectomía bilateral es la de mayor ganancia con 13 años mientras que la cirugía reductora de riesgo en trompas y ovarios gana 4,95 años con respecto al grupo de seguimiento. Conclusiones Las tres opciones evaluadas son aceptables, pero de ellas la de mayor ganancia en la supervivencia es la combinación de cirugía reductora de riesgo en trompas y ovarios con mastectomía bilateral.(AU)

ABSTRACT Objective To assess the usefulness of risk reduction salpingo-oophorectomy in cases with mutation of the BRCA1 gene in Colombia. Material and Methods Cost-effectiveness analysis in which three processes are incorporated: a. Patients with screening tests for breast and ovarian cancer. b. Risk reduction surgery in the fallopian tubes and ovaries c. Reductive surgery in the fallopian tubes and ovaries with bilateral mastectomy. The outcome is evaluated as the gain in years of survival. Results The cohort with risk reduction surgery in the fallopian tubes and ovaries and bilateral mastectomy is the one with the highest gain with 13 years, while the risk reduction surgery in the fallopian tubes and ovaries gain 4.95 years with respect to the follow-up group. Conclusions The three options evaluated are acceptable, but of them the one with the greatest gain in survival is the combination of risk-reducing surgery in the fallopian tubes and ovaries with bilateral mastectomy.(AU)

Humans , Female , Ovarian Neoplasms/surgery , Breast Neoplasms/surgery , Ovariectomy/rehabilitation , Genes, BRCA1 , Preventive Medicine , Markov Chains , Colombia , Cost-Effectiveness Evaluation
Journal of Breast Cancer ; : 297-305, 2018.
Article in English | WPRIM | ID: wpr-716746


PURPOSE: The characteristic expression of DNA damage response proteins in familial breast cancers with BRCA1, BRCA2, or non-BRCA1/2 mutations has not been analyzed in Chinese patients. Our study aimed to assess the differential expression of microcephalin 1 (BRIT1), ATM serine/threonine kinase (ATM), checkpoint kinase 2 (CHEK2), BRCA1, RAD51 recombinase (RAD51), and poly (ADP-ribose) polymerase 1 (PARP-1) and establish the profile of Chinese familial breast cancers with different mutation status. METHODS: We constructed five tissue microarrays from 183 familial breast cancer patients (31 with BRCA1 mutations; 14 with BRCA2 mutations, and 138 with non-BRCA1/2 mutations). The DNA response and repair markers used for immunohistochemistry analysis included BRIT1, ATM, CHEK2, BRCA1, RAD51, and PARP-1. The expressions of these proteins were analyzed in BRCA1/2 mutated tumors. The association between pathologic characteristics with BRCA1/2 mutation status was also analyzed. RESULTS: In familial breast cancer patients, BRCA1 mutated tumors were more frequent with high nuclear grade, estrogen receptor/progesterone receptor/human epidermal growth factor receptor 2 negative, low Ki-67, and positive CK5/6. BRCA1 mutated tumors had lower CHEK2 and higher cytoplasmic BRIT1 expression than BRCA2 and non-BRCA1/2 mutation tumors. BRCA2-associated tumors showed higher CHEK2 and cytoplasmic RAD51 expression than those in other groups. Nuclear PARP-1 expression in BRCA1/2-associated tumors was significantly higher than in non-BRCA1/2 mutation tumors. Moreover, we found quite a few of negative PARP-1 expression cases in BRCA1/2 mutated groups. CONCLUSION: The clinicopathologic findings of BRCA1-associated Chinese familial breast cancers were similar to the results of other studies. Chinese familial breast cancer patients with BRCA1/2 mutations might have distinctive expression of different DNA damage response proteins. The reduced expression of PARP-1 in Chinese BRCA1/2 mutated breast cancer patients could influence the therapeutic outcome of PARP-1 inhibitors.

Asians , Breast Neoplasms , Breast , Checkpoint Kinase 2 , Cytoplasm , DNA Damage , DNA Repair , DNA , Estrogens , Genes, BRCA1 , Genes, BRCA2 , Humans , Immunohistochemistry , Phosphotransferases , Rad51 Recombinase , ErbB Receptors
Article in English | WPRIM | ID: wpr-716093


OBJECTIVE: The aim of the present study was to assess the frequency of germline mutations in patients with peritoneal carcinoma (PC) or the fallopian tube carcinoma (FTC), using a multi-gene panel. METHODS: Twenty-six patients diagnosed with either PC or FTC between January 2013 and December 2016 were recruited consecutively. Germline DNA was sequenced using a 6-gene next generation sequencing (NGS) panel following genetic counseling. Surgico-medical information was obtained from hospital records. Genetic variations were detected using the panel and were cross-validated by Sanger direct sequencing. RESULTS: Germline BRCA1/2 mutations were identified in 6 patients (23.1%). Four were detected in patients with PC and 2 were in FTC patients. No mutations were detected in TP53, PTEN, CDH1, or PALB2. We identified 11 variant of uncertain significance (VUS) in 9 patients; 2 in BRCA1, 3 in BRCA2, 2 in TP53, and 4 in CDH1. We also detected a CDH1 c.2164+16->A VUS in 3 patients. CONCLUSION: The prevalence of germline BRCA1/2 mutations in patients with PC or FTC is comparable to that of BRCA1/2 mutations in epithelial ovarian cancer patients.

DNA , Fallopian Tube Neoplasms , Fallopian Tubes , Female , Genes, BRCA1 , Genes, BRCA2 , Genetic Counseling , Genetic Variation , Germ-Line Mutation , Hospital Records , Humans , Ovarian Neoplasms , Peritoneal Neoplasms , Peritoneum , Prevalence
IBJ-Iranian Biomedical Journal. 2018; 22 (3): 210-214
in English | IMEMR | ID: emr-192470


Background: Previous studies have suggested that BRCA1 dysregulation has been shown to have a role in triplenegative phenotypic manifestation. However, differences of BRCA1 expression, as a tumor suppressor gene, have rarely been investigated between luminal and triple-negative breast tumors. Therefore, the present study attempted to compare the BRCA1 expression in triple-negative with luminal breast tumors and its association with the clinicopathologic characteristics of patients

Methods: BRCA1 expression was evaluated by real-time PCR in 26 triple-negative and 27 luminal breast tumors

Results: The results revealed that there is a high frequency of BRCA1 underexpression in both triple-negative and luminal breast tumors. The BRCA1 underexpression was related to young age at diagnosis, lymph node involvement, and grade III tumors

Conclusion: The observations suggest that decreased BRCA1 expression, regardless of tumor subtype, has a general role in breast malignancy and associated with poor prognostic features in breast tumors

Humans , Female , Breast Neoplasms/diagnosis , Genes, BRCA1 , Triple Negative Breast Neoplasms , Real-Time Polymerase Chain Reaction
Biol. Res ; 51: 58, 2018. graf
Article in English | LILACS | ID: biblio-1011402


BACKGROUND: Studies have demonstrated that transforming growth factor beta-1 (TGF-ß1) exhibits oncogenic activity in different types of cancer, including ovarian cancer (OC). However, its regulatory mechanism in OC and whether TGF-ß1 is involved in chemosensitivity regulation remains unclear. Thus, the aim of this study was to investigate the role of TGF-ß1 in OC. METHODS: The OC cell line SKOV3 was employed, and TGF-ß1 overexpression or knockdown vectors were constructed. The cell proliferation of SKOV3 was evaluated with the cell counting kit (CCK8) kit after treatment with different concentrations of cis-platinum. Western blot and protein immunoprecipitation were employed to detect changes in BRCA1 and Smad3 expression and their interactions. Tumor growth in nude mice was evaluated. RESULTS: The results showed that TGF-ß1 knockdown increased chemosensitivity by promoting BRCA1 expression and Smad3 phosphorylation. In vivo studies showed that TGF-ß1 knockdown significantly inhibited the growth of tumors, also by upregulating BRCA1 expression and Smad3 phosphorylation. CONCLUSION: Taken together, our results suggest that TGF-ß1 knockdown inhibits tumor growth and increases chemosensitivity by promotion of BRCA1/Smad3 signaling.

Humans , Animals , Male , Female , Ovarian Neoplasms/metabolism , Down-Regulation/physiology , Genes, BRCA1/physiology , Smad3 Protein/physiology , Transforming Growth Factor beta1/physiology , Ovarian Neoplasms/pathology , Ovarian Neoplasms/drug therapy , Immunohistochemistry , Cells, Cultured , Blotting, Western , Drug Resistance, Neoplasm/physiology , Tumor Suppressor Proteins/physiology , Cell Line, Tumor , Cell Proliferation , Smad3 Protein/analysis , Transforming Growth Factor beta1/analysis , Gene Knockdown Techniques , Real-Time Polymerase Chain Reaction , Mice, Inbred BALB C
Rev. saúde pública (Online) ; 52: 94, 2018. tab, graf
Article in English | LILACS | ID: biblio-979019


ABSTRACT OBJECTIVE: To analyze the cost effectiveness of the diagnostic program for the germline mutation in BRCA1/2 genes and of preventative strategies for the relatives of patients diagnosed with ovarian cancer associated with this mutation. METHODS: The study analyzed the cost effectiveness by developing an analysis of the Markov decision process from the perspective of the National Health System. The strategies compared reflect upon the adoption of genetic testing and preventative strategies for relatives or the usual care currently proposed. The incremental cost-effectiveness ratio was expressed in terms of cost per case avoided. The sensitivity analysis was performed in a univariate and deterministic manner. RESULTS: The study showed increments for effectiveness and for costs when performing genetic testing and adopting prophylactic measures for family members. The incremental cost-effectiveness ratio was estimated at R$908.58 per case of cancer avoided, a figure considered lower than the study's cost-effectiveness threshold (R$7,543.50). CONCLUSIONS: The program analyzed should be considered a cost-effective strategy for the national situation. Studies in various other countries have reached similar conclusions. One possible ramification of this research might the need to perform a budgetary-impact analysis of making the program one of the country's health policies.

Humans , Female , Adolescent , Adult , Ovarian Neoplasms/genetics , Ovarian Neoplasms/prevention & control , Program Evaluation/economics , Germ-Line Mutation/genetics , Genes, BRCA1 , Genes, BRCA2 , Ovarian Neoplasms/economics , Reference Values , Brazil , Breast Neoplasms/genetics , Genetic Testing/economics , Reproducibility of Results , Risk Factors , Markov Chains , Cost-Benefit Analysis , Middle Aged
Rev. méd. Urug ; 33(2): 102-107, Jun. 2017.
Article in Spanish | LILACS | ID: biblio-859973


Introducción: el cáncer de mama representa la primera causa de muerte por cáncer en mujeres de Uruguay. Se estima que cerca de 7% son causados por mutaciones en el ácido desoxirribonucleico germinal. Los costos de la secuenciación genética han descendido dramáticamente gracias a la aparición de la secuenciación de nueva generación (NGS). El cambio tecnológico abrió una nueva etapa en el estudio del cáncer hereditario en nuestro país. Objetivo: comunicar los resultados de la utilización de tecnología NGS y paneles multigénicos en familias uruguayas con alto riesgo de cáncer de mama hereditario. Pacientes y método: se secuenciaron 135 familias de alto riesgo que provenían de la consulta de consejería genética que funciona en el Grupo Colaborativo Uruguayo: Investigación de afecciones oncológicas hereditarias. Cuando la historia familiar sugería claramente un síndrome de cáncer de mama y ovario hereditario se efectuó secuenciación NGS exclusiva de los genes BRCA1 y 2; cuando el patrón familiar no configuraba claramente se utilizó un panel multigénico. Resultados: se efectuó NGS exclusiva de genes BRCA1 y 2 en 62 familias y un panel multigénico en 73 familias. Se identificaron en total 29 mutaciones patógenas (21 en genes BRCA y 8 en otros genes). Dos de ellas fueron noveles y tres pueden considerarse recurrentes en la población uruguaya. Conclusiones: este trabajo es el primero en Uruguay en reportar el resultado de esta nueva tecnología en el cáncer de mama hereditario. El hallazgo de 29 mutaciones patógenas nos ayuda a delinear el perfil mutacional de nuestro país.

Introduction: breast cancer is women's first cause of death in Uruguay. According to estimations, around 7% of cases result from germinal mutations by deoxyribonucleic acid. The cost of genetic sequencing has dramatically dropped thanks to the arrival of next-generation sequencing (NGS). This technological change opened a new era in the study of hereditary cancer in our country. Objective:to communicate the results of using NGS technology and multigenic panels in Uruguayan families with high risk of hereditary breast cancer. Method: 135 high risk families referred by the genetic counselling consultation that is provided at the Uruguayan Collaborative Group (Hereditary Oncological Conditions Research) were sequenced. When the family history clearly suggested hereditary breast and ovary cancer was a possibility, exclusive NGS sequencing was done for BRCA1 and BRCA2 genes; when the family pattern was not clear to this respect, multigenic panels were used. Results: exclusive NGS sequencing for BRCA1 and BRCA2 genes was done in 62 families, and multigenic panels were used in 73 families. 29 pathogenic mutations were identified (21 in BRCA genes and 8 in other genes). Two of them were new to the disease and three could be considered recurrent in the Uruguayan population. Conclusions:this study is the first one in Uruguay to report the results of this new technology in hereditary breast cancer. The finding of 29 pathogenic mutations contributes to outlining the mutational profile of our country.

Introdução: o câncer de mama é a primeira causa de morte por câncer em mulheres no Uruguai. Estima-se que aproximadamente 7% sejam causados por mutações no ácido desoxirribonucleico germinal. Os custos da sequenciação genética diminuíram dramaticamente graças ao aparecimento da sequenciação de nova geração (NGS). Esta nova tecnologia deu inicio a uma nueva etapa no estudo do câncer hereditário no nosso país. Objetivo: comunicar os resultados da utilização de tecnologia NGS e painéis mutagênicos em famílias uruguaias com alto risco de câncer de mama hereditário. Pacientes e método: 135 famílias de alto risco originárias do aconselhamento genético que funciona no Grupo Colaborativo Uruguaio: Pesquisa de afecções oncológicas hereditárias foram sequenciadas. Quando a história familiar sugeria uma síndrome de câncer de mama e ovário hereditários fez-se a secuenciacao NGS exclusivamente dos genes BRCA1 e 2; quando o padrão familiar não era claro foi utilizado um painel multigênico. Resultados: realizou-se NGS exclusivamente de genes BRCA1 e 2 em 62 famílias e um painel multigênico em 73 famílias. Foram identificadas 29 mutações patogênicas (21 em genes BRCA e 8 em outros genes). Duas eram novas e três podem ser consideradas como recorrentes na população uruguaia. Conclusões: este trabalho é o primeiro que apresenta os resultados desta nova tecnologia aplicada ao câncer de mama hereditário no Uruguai. O achado de 29 mutações patogênicas ajuda a definir o perfil mutacional do nosso país.

Breast Neoplasms/genetics , Genes, BRCA1 , Genes, BRCA2
Article in English | WPRIM | ID: wpr-167305


PURPOSE: Unclassified variants (UVs) of BRCA1 and BRCA2 genes are not defined as pathogenic for breast cancer, and their clinical significance currently remains undefined. Therefore, this study was conducted to identify potentially pathogenic UVs by comparing their prevalence between breast cancer patients and controls. MATERIALS AND METHODS: A total of 328 breast cancer patients underwent BRCA1/2 genetic screening at the National Cancer Center of Korea. Genetic variants of BRCA genes that were categorized as unclassified according to the Breast Cancer Information Core database were selected based on allelic frequency, after which candidate variants were genotyped in 421 healthy controls. We also examined family members of the study participants. Finally, the effects of amino acid substitutions on protein structure and function were predicted in silico. RESULTS: Genetic tests revealed 33 UVs in BRCA1 and 47 in BRCA2. Among 15 candidates genotyped in healthy controls, c.5339T>C in BRCA1 and c.6029T>G, c.7522G>A in BRCA2 were not detected. Moreover, the c.5339T>C variant in the BRCA1 gene was detected in four patients with a family history of breast cancer. This nonsynonymous variant (Leu1780Pro) in the BRCA1 C-terminal domain was predicted to have an effect on BRCA1 protein structure/function. CONCLUSION: This study showed that comparison of genotype frequency between cases and controls could help identify UVs of BRCA genes that are potentially pathogenic. Moreover, ourfindings suggest that c.5339T>C in BRCA1 might be a pathogenic variant for patients and their families.

Amino Acid Substitution , BRCA1 Protein , Breast Neoplasms , Breast , Computer Simulation , Genes, BRCA1 , Genes, BRCA2 , Genetic Testing , Genotype , Humans , Korea , Prevalence
Article in English | WPRIM | ID: wpr-101939


PURPOSE: Familial pancreatic cancer describes families with at least two first-degree relatives with pancreatic cancer that do not fulfil the criteria of other inherited tumor syndromes with increased risks of pancreatic cancer. Although much has been learned regarding the aggregation of pancreatic cancer in some families, the genetic basis for this familial aggregation is poorly understood. This study evaluated the prevalence of 10 Polish founder mutations in four genes among individuals from families with diagnosed familial pancreatic cancer syndrome and assessed their possible association with the familial pancreatic cancer (FPC) risk in Poland. MATERIALS AND METHODS: In this study, 400 FPC individuals and 4,000 control subjects were genotyped for founder mutations in BRCA1 (5382insC, 4153delA, C61G), CHEK2 (1100delC, IVS2+1G>A, del5395, I157T), NBS1 (657del5), and PALB2 (509_510delGA, 172_175delTTGT) genes. RESULTS: A statistically significant association was observed between the 172_175delTTGT mutation of the PALB2 gene and an increased risk of FPC syndrome (odds ratio [OR], 10.05; p=0.048). In addition, an increased risk of cancer was observed in the FPC family members with a BRCA1 mutation (OR, 6.72; p=0.006). Novel associations were found between the FPC family members with cancer and CHEK2 mutations (OR, 2.26; p=0.008) with a noticeable contribution of the missense variant, I157T of CHEK2 (OR, 2.17; p=0.026). CONCLUSION: The founder mutations in the genes, BRCA1, PALB2, and CHEK2, cause a small percentage of familial pancreatic cancer syndrome in the Polish population. Following confirmation in larger studies, these mutations can be added to the panel of genes to be tested in families with a diagnosis of FPC syndrome.

Diagnosis , Genes, BRCA1 , Humans , Pancreatic Neoplasms , Poland , Prevalence