Asunto(s)
Quimioradioterapia Adyuvante/métodos , Neoplasias Endometriales/terapia , Inestabilidad de Microsatélites/efectos de la radiación , Anciano , Anciano de 80 o más Años , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Procedimientos Quirúrgicos de Citorreducción , Neoplasias Endometriales/patología , Femenino , Humanos , Estimación de Kaplan-Meier , Persona de Mediana Edad , Recurrencia Local de Neoplasia , Estudios Retrospectivos , Resultado del TratamientoAsunto(s)
Neoplasias del Colon/patología , Neoplasias del Colon/cirugía , Manejo de Especímenes/métodos , Manejo de Especímenes/normas , Quimioradioterapia , Neoplasias del Colon/tratamiento farmacológico , Neoplasias del Colon/genética , Neoplasias del Colon/radioterapia , Humanos , Inestabilidad de Microsatélites/efectos de los fármacos , Inestabilidad de Microsatélites/efectos de la radiación , Mutación , Invasividad Neoplásica , Estadificación de Neoplasias/métodos , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas B-raf/genética , Proteínas Proto-Oncogénicas p21(ras) , Inducción de Remisión , Proteínas ras/genéticaRESUMEN
PURPOSE: To investigate microsatellite instability (MSI) in radiation-induced murine tumours, its dependence on tissue (haemopoietic, intestinal, mammary, brain and skin) and radiation type. MATERIALS AND METHODS: DNA from spontaneous, X-ray or neutron-induced mouse tumours were used in Polymerase Chain Reactions (PCR) with mono- or di-nucleotide repeat markers. Deviations from expected allele size caused by insertion/deletion events were assessed by capillary electrophoresis. RESULTS: Tumours showing MSI increased from 16% in spontaneously arising tumours to 23% (P = 0.014) in X-ray-induced tumours and rising again to 83% (P << 0.001) in neutron-induced tumours. X-ray-induced Acute Myeloid Leukaemias (AML) had a higher level of mono-nucleotide instability (45%) than di-nucleotide instability (37%). Fifty percent of neutron-induced tumours were classified as MSI-high for mono-nucleotide markers and 10% for di-nucleotide markers. Distribution of MSI varied in the different tumour types and did not appear random. CONCLUSIONS: Exposure to ionising radiation, especially neutrons, promotes the development of MSI in mouse tumours. MSI may therefore play a role in mouse radiation tumourigenesis, particularly following high Linear Energy Transfer (LET) exposures. MSI events, for a comparable panel of genome-wide markers in different tissue types, were not randomly distributed throughout the genome.
Asunto(s)
Inestabilidad de Microsatélites/efectos de la radiación , Repeticiones de Microsatélite/efectos de la radiación , Neoplasias Inducidas por Radiación/etiología , Neoplasias Inducidas por Radiación/genética , Neutrones/efectos adversos , Rayos X/efectos adversos , Animales , Línea Celular Tumoral , ADN/aislamiento & purificación , ADN/metabolismo , ADN/efectos de la radiación , Electroforesis Capilar , Fibroblastos/patología , Leucemia Mieloide Aguda/patología , Ratones , Ratones Endogámicos C57BL , Proteína 2 Homóloga a MutS/metabolismo , Neoplasias Inducidas por Radiación/clasificación , Neoplasias Inducidas por Radiación/patología , Reacción en Cadena de la PolimerasaRESUMEN
Deficiencies in individual DNA repair systems are involved in both de novo and therapy-related acute myeloid leukaemia (t-AML), as indicated by genetic markers involving nucleotide excision repair (NER gene polymorphisms), double-strand-break (DSB) or mismatch repair (microsatellite instability (MSI)). We modified a host cell reactivation (HCR) assay for functional DNA repair system analysis of living primary haematopoietic cells; 2 x 10(5) normal peripheral blood lymphocytes (PBLs) and cord blood CD34+ progenitor cells were cryopreserved, thawed and transfected with 75-250 ng luciferase reporter plasmid (pCMVLuc) using DEAE-dextran (0.1 mg/mL) in a transfection volume of 250 microL. We obtained luciferase activities of approximately 300-fold above background in CD34+ progenitor cells and approximately 2000-fold in PBLs, thus rendering these cells applicable for DNA repair analysis. We then evaluated the NER (UV-irradiated pCMVLuc) and DSB repair capacity (linearized pCMVLuc) of normal lymphocytes and several leukaemic cell lineages. Kasumi-1 and HL-60 AML cells exhibited a reduced NER capacity compared to normal GM03715 lymphocytes, PBLs and CD34+ progenitor cells (6.2 +/- 0.9%, 6.5 +/- 0.9% vs. 12.3 +/- 1.8%, 13.5 +/- 0.7% and 13.5 +/- 2.0%, respectively). Kasumi-1 AML tells exhibited a reduced DSB repair capacity compared to AG10107 and GM03715 normal lymphocytes as well as CEM acute T-cell lymphoblastic leukaemia cells (6.4 +/- 0.8% vs. 10.8 +/- 0.7%, 27.3 +/- 1.1% and 20.5 +/- 1.6%, respectively). The modified HCR assay can be used for functional DNA repair analysis in living cells of patients with pre- and post-leukaemic conditions as well as in leukaemic blasts to elucidate the role of DNA repair in de novo and t-AML leukaemogenesis and to determine the individual susceptibility to t-AML prior to chemotherapy.
Asunto(s)
Reparación del ADN , ADN/genética , Genes Reporteros/genética , Células Madre Hematopoyéticas/metabolismo , Luciferasas/genética , Luciferasas/metabolismo , Linfocitos/metabolismo , Células Cultivadas , ADN/biosíntesis , Roturas del ADN de Doble Cadena/efectos de la radiación , Daño del ADN , Activación Enzimática , Femenino , Técnicas Genéticas , Células HL-60 , Humanos , Leucemia Monocítica Aguda/genética , Leucemia-Linfoma de Células T del Adulto/genética , Luciferasas/análisis , Linfocitos/clasificación , Inestabilidad de Microsatélites/efectos de la radiación , Plásmidos/genética , Traumatismos por Radiación/genética , Transfección , Rayos Ultravioleta/efectos adversosRESUMEN
The effects of X-ray and carbon ion irradiation on DNA and genes in head and neck carcinoma cells were examined. Four head and neck cancer cell lines (squamous cell carcinoma, salivary gland cancer, malignant melanoma, normal keratinocyte) were treated with 1, 4, and 7 GyE of carbon ion, or 1, 4, and 8 Gy of X-ray, respectively. DNA and RNA in the treated cells were extracted and purified. PCR-LOH (polymerase chain reaction-loss of heterozygosity) analysis with 6 microsatellite regions on chromosome 17 was performed to determine DNA structural damage, and then microarray analysis was performed to reveal changes in gene expression. PCR-LOH analysis detected high LOH in cells treated by radiation, indicating that most of the damage by X-ray occurred in the target region on one of the homologous chromosomes. However, carbon ion caused homo-deletion, which means deletion of the counterparts in both homologous chromosomes.