RESUMO
VHL is mutated in the majority of patients with clear cell renal cell carcinoma (ccRCC), with conflicting clinical relevance. Recent studies have identified recurrent mutations in histone modifying and chromatin remodeling genes, including BAP1, PBRM1, SETD2, KDM6A, and JARID1c. Current evidence suggests that BAP1 mutations are associated with aggressive disease. The clinical significance of the remaining genes is unknown. In this study, targeted sequencing of VHL and JARID1c (entire genes) and coding regions of BAP1, PBRM1, SETD2, and KDM6A was performed on 132 ccRCCs and matched normal tissues. Associations between mutations and clinical and pathological outcomes were interrogated. Inactivation of VHL (coding mutation or promoter methylation) was seen in 75% of ccRCCs. Somatic noncoding VHL alterations were identified in 29% of ccRCCs and may be associated with improved overall survival. BAP1 (11%), PBRM1 (33%), SETD2 (16%), JARID1c (4%), and KDM6A (3%) mutations were identified. BAP1-mutated tumors were associated with metastatic disease at presentation (P = 0.023), advanced clinical stage (P = 0.042) and a trend towards shorter recurrence free survival (P = 0.059) when compared with tumors exclusively mutated for PBRM1. Our results support those of recent publications pointing towards a role for BAP1 and PBRM1 mutations in risk stratifying ccRCCs. Further investigation of noncoding alterations in VHL is warranted.
Assuntos
Carcinoma de Células Renais/genética , Proteínas de Transporte/genética , Neoplasias Renais/genética , Proteínas Nucleares/genética , Fatores de Transcrição/genética , Proteínas Supressoras de Tumor/genética , Ubiquitina Tiolesterase/genética , Idoso , Carcinoma de Células Renais/patologia , Montagem e Desmontagem da Cromatina , Proteínas do Citoesqueleto , Metilação de DNA/genética , Proteínas de Ligação a DNA , Feminino , Histona Desmetilases/genética , Histona-Lisina N-Metiltransferase/genética , Humanos , Neoplasias Renais/patologia , Masculino , Pessoa de Meia-Idade , Chaperonas Moleculares , Oxirredutases N-Desmetilantes/genética , Estudos RetrospectivosRESUMO
BACKGROUND: The diagnosis and treatment of prostate cancer is a challenging global healthcare issue requiring significant molecular research. Such research frequently utilizes fresh frozen human tissue which needs to be obtained in a manner acceptable to the pathologist which does not compromise tumor diagnosis or staging. METHODS: Radical prostatectomy specimens were handled in a standardized method before being sliced fresh. Leaving the margins intact, multiple cylindrical cores were removed using a large skin punch and the sites were marked on a prostate map. The cylindrical cores were placed onto individual, pre-numbered foil squares and snap frozen in liquid nitrogen. Prostate maps were aligned with formalin-fixed paraffin embedded hematoxylin and eosin stained sections of the sampled slice to select tumor regions. Frozen tumor tissue cylinders were processed taking one section for hematoxylin and eosin staining, 6 µm × 50 µm sections for molecular studies and a further section for hematoxylin and eosin staining. This was performed for the length of the cylinder. RESULTS: A total of 150 prostates have been removed and sliced using this technique. Pathological assessment remained uncompromised. Using the sequential hematoxylin and eosin stained frozen sections, cellularity could be monitored closely in tissues processed for research. The yield of RNA and DNA extracted was high (tumor mean 2.4 µg (RNA) and 12.7 ng per 300 µm tissue) and of high quality (mean tumor RIN 5.9). CONCLUSIONS: This novel, rapid sampling and processing method provides high quality tissue for research without compromising pathology.