RESUMEN
PURPOSE: The molecular mechanism of breast and/or ovarian cancer susceptibility remains unclear in the majority of patients. While germline mutations in the regulatory non-coding regions of BRCA1 and BRCA2 genes have been described, screening has generally been limited to coding regions. The aim of this study was to evaluate the contribution of BRCA1/2 non-coding variants. METHODS: Four BRCA1/2 non-coding regions were screened using high-resolution melting analysis/Sanger sequencing or next-generation sequencing on DNA extracted from index cases with breast and ovarian cancer predisposition (3926 for BRCA1 and 3910 for BRCA2). The impact of a set of variants on BRCA1/2 gene regulation was evaluated by site-directed mutagenesis, transfection, followed by Luciferase gene reporter assay. RESULTS: We identified a total of 117 variants and tested twelve BRCA1 and 8 BRCA2 variants mapping to promoter and intronic regions. We highlighted two neighboring BRCA1 promoter variants (c.-130del; c.-125C > T) and one BRCA2 promoter variants (c.-296C > T) inhibiting significantly the promoter activity. In the functional assays, a regulating region within the intron 12 was found with the same enhancing impact as within the intron 2. Furthermore, the variants c.81-3980A > G and c.4186-2022C > T suppress the positive effect of the introns 2 and 12, respectively, on the BRCA1 promoter activity. We also found some variants inducing the promoter activities. CONCLUSION: In this study, we highlighted some variants among many, modulating negatively the promoter activity of BRCA1 or 2 and thus having a potential impact on the risk of developing cancer. This selection makes it possible to conduct future validation studies on a limited number of variants.
Asunto(s)
Proteína BRCA1/genética , Proteína BRCA2/genética , Genes BRCA1 , Genes BRCA2 , Síndrome de Cáncer de Mama y Ovario Hereditario/genética , Adulto , Anciano , Estudios de Cohortes , Biología Computacional , Femenino , Predisposición Genética a la Enfermedad , Mutación de Línea Germinal , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Intrones/genética , Persona de Mediana Edad , Linaje , Polimorfismo de Nucleótido Simple , Regiones Promotoras Genéticas/genética , Regiones no Traducidas/genéticaRESUMEN
The earthquake in Haiti in January 2010 resulted in more than 220,000 deaths and over 300,000 injured and was one of the greatest mass casualties in recent history. "Doctors Without Borders" started a medical relief response immediately after the earthquake, building up to the biggest disaster relief activity in the organization's history. Roughly 173,000 medical consultations and more than 11,700 surgical interventions were performed in 26 medical facilities during the first 4 months. A particular challenge was the sheer number of patients in a situation with a completely destroyed medical infrastructure. While the initial phase mainly focused on life saving surgery, the second phase concentrated on reconstructive surgery of the extremities. Crucial for effective patient care is an ability to act early and employ surgical techniques which are adapted to the overall situation. The following article is a personal report of the early emergency response from the viewpoint of two orthopedic trauma surgeons, who have surgical careers in Germany and also frequently volunteer for "Doctors Without Borders".