RESUMO
BACKGROUND: The majority of patients with stage III-IV epithelial ovarian cancer (EOC) relapse after initially responding to platinum-based chemotherapy, and develop resistance. The genomic features involved in drug resistance are unknown. To unravel some of these features, we investigated the mutational profile of genes involved in pathways related to drug sensitivity in a cohort of matched tumors obtained at first surgery (Ft-S) and second surgery (Sd-S). PATIENTS AND METHODS: Matched biopsies (33) taken at Ft-S and Sd-S were selected from the 'Pandora' tumor tissue collection. DNA libraries for 65 genes were generated using the TruSeq Custom Amplicon kit and sequenced on MiSeq (Illumina). Data were analyzed using a high-performance cluster computing platform (Cloud4CARE project) and independently validated. RESULTS: A total of 2270 somatic mutations were identified (89.85% base substitutions 8.19% indels, and 1.92% unknown). Homologous recombination (HR) genes and TP53 were mutated in the majority of Ft-S, while ATM, ATR, TOP2A and TOP2B were mutated in the entire dataset. Only 2% of mutations were conserved between matched Ft-S and Sd-S. Mutations detected at second surgery clustered patients in two groups characterized by different mutational profiles in genes associated with HR, PI3K, miRNA biogenesis and signal transduction. CONCLUSIONS: There was a low level of concordance between Ft-S and Sd-S in terms of mutations in genes involved in key processes of tumor growth and drug resistance. This result suggests the importance of future longitudinal analyses to improve the clinical management of relapsed EOC.
Assuntos
Adenocarcinoma de Células Claras/genética , Adenocarcinoma Mucinoso/genética , Cistadenocarcinoma Seroso/genética , Neoplasias do Endométrio/genética , Genes Neoplásicos/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Mutação/genética , Neoplasias Ovarianas/genética , Adenocarcinoma de Células Claras/mortalidade , Adenocarcinoma de Células Claras/secundário , Adenocarcinoma de Células Claras/terapia , Adenocarcinoma Mucinoso/mortalidade , Adenocarcinoma Mucinoso/secundário , Adenocarcinoma Mucinoso/terapia , Adulto , Idoso , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Biópsia , Terapia Combinada , Cistadenocarcinoma Seroso/mortalidade , Cistadenocarcinoma Seroso/secundário , Cistadenocarcinoma Seroso/terapia , Resistencia a Medicamentos Antineoplásicos/genética , Neoplasias do Endométrio/mortalidade , Neoplasias do Endométrio/secundário , Neoplasias do Endométrio/terapia , Feminino , Seguimentos , Recombinação Homóloga , Humanos , Estudos Longitudinais , Metástase Linfática , Pessoa de Meia-Idade , Gradação de Tumores , Recidiva Local de Neoplasia/genética , Recidiva Local de Neoplasia/mortalidade , Recidiva Local de Neoplasia/patologia , Recidiva Local de Neoplasia/terapia , Estadiamento de Neoplasias , Neoplasias Ovarianas/mortalidade , Neoplasias Ovarianas/patologia , Neoplasias Ovarianas/terapia , Prognóstico , Estudos Retrospectivos , Taxa de SobrevidaRESUMO
The content of chlorides, nitrites, nitrates, phosphates and sulphates was used to classify 45 donkey's milk samples collected from different Italian regions. A method employing ion exchange chromatography with conductivity detector and chemical suppression was used. The quantitative results indicated phosphates (569.4-1304.4 mg kg(-1)) and chlorides (545.9-1757.9 mg kg(-1)) as being the most abundant anions, followed by sulphates (109.5-200.7 mg kg(-1)). The concentrations of nitrites and nitrates were found to be lower at 5.6 and 5.5 mg kg(-1) respectively. The data set was subdivided into three groups according to the region of origin of milk, and was statistically evaluated by analysis of variance (ANOVA). Concentrations of chlorides and nitrites showed a significant difference among farms (p < 0.001). In a first discriminant analysis procedure, functions based on linear combinations of the log(e)-transformed element concentrations of anions were generated to classify donkey's milk samples from different regions. In an alternative approach, a three-step discriminant analysis procedure to classify a milk sample was tested. The results obtained led to a correct classification of donkey's milk samples based on their anions content with 91-98% of the samples being correctly classified. The procedure proved to be very simple, so it could be used as an evaluation method for the traceability of donkey's milk, thus defending this unique product against fraud or commercial disputes.
