RESUMEN
Sensitivity to ultraviolet radiation was studied in lymphoblastoid cell lines derived from 32 members of two families with histories of multiple primary melanomas in several generations. As assayed by colony formation in agar or by trypan blue exclusion following irradiation, cellular sensitivity showed a bimodal distribution. All persons with melanoma or multiple moles were in the sensitive group, while some family members exhibited responses similar to those of controls. Cells from four cases of sporadic melanoma showed normal levels of sensitivity. The data are consistent with a dominantly inherited ultraviolet light sensitivity associated with these examples of familial melanoma. Spontaneous and ultraviolet light-induced sister chromatid exchange frequencies were similar to those in control cell lines. No defect in excision repair was detected in any of the above cell lines, but the sensitive group showed postirradiation inhibition of DNA replication intermediate between controls and an excision-deficient xeroderma pigmentosum cell line.
Asunto(s)
Linfocitos/efectos de la radiación , Melanoma/genética , Neoplasias Cutáneas/genética , Rayos Ultravioleta , Supervivencia Celular/efectos de la radiación , Células Cultivadas , ADN de Neoplasias/biosíntesis , Humanos , Linfocitos/metabolismo , Melanoma/metabolismo , Melanoma/fisiopatología , Neoplasias Cutáneas/metabolismo , Neoplasias Cutáneas/fisiopatologíaRESUMEN
Ionizing radiation sensitive, mutant human lymphoblastoid cell lines derived from patients with Huntington's disease (HD), or ataxia telangiectasia (AT) both showed cross sensitivity to bleomycin, as assayed by reduced cell viability and increased frequency of chromosome aberrations compared to normal controls. In contrast to AT cells which failed to show inhibition of DNA synthesis after exposure to ionizing radiation, or bleomycin treatment, the sensitive cells from HD patients had depressed rates of DNA synthesis after damage with these agents, similar to that seen in normal cells. In terms of progression through the cell cycle bleomycin damaged AT cells moved from G1 into S and from S to G2 + M at almost the same rate as untreated cells. Bleomycin treated HD cells showed a large proportion of cells blocked in G1, cells were slowed down in S, the rate of entry to G2 + M was reduced and only 5% of cycling cells reached G2. Progress through the cell cycle in normal cells exposed to bleomycin showed a partial block in G1 and the rate of entry to G2 + M was reduced. These differences in response of normal, AT and HD cells to ionizing radiation and bleomycin treatment indicates that the defect underlying the sensitivity is different in HD cells from that in AT cells.
Asunto(s)
Ataxia Telangiectasia/sangre , Bleomicina/farmacología , Enfermedad de Huntington/sangre , Linfocitos/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Aberraciones Cromosómicas , Replicación del ADN/efectos de los fármacos , Humanos , Linfocitos/efectos de la radiaciónRESUMEN
Three strains of Salmonella typhimurium carrying frameshift mutations affectin g the histidine genes (hisC3076, hisD3052 and hisC207) showed increased sensitivity to mutagenesis by ICR-191 (as judged by measuring back mutation to prototrophy), if they were made deficient in excision repair by deleting the uvrB gene. One frameshift strain, hisC3076, also showed increased sensitivity to mutagenesis by ICR-191 when it carried either of two different polA alleles, whereas the hisD3052 and hisC207 frameshifts reduced sensitivity to mutagenesis in the presence of these alleles. Studies of spontaneous back mutation to prototrophy revealed significant mutator effects of the polA1 mutation on reversion of the hisD3052 frameshift and of the polA3 mutation on reversion of the hisC3076 frameshift. Other smaller mutator effects of the polA alleles on complex interactions between different polA alleles and different frameshift mutations, it is tentatively suggested that deletion frameshifts may arise mainly during DNA replication, while addition frameshifts may arise mainly during post-replication repair.
Asunto(s)
Mutación , Salmonella typhimurium/metabolismo , Alelos , Reparación del ADN , Replicación del ADN , Código Genético , Genotipo , Histidina/metabolismo , Transcripción GenéticaRESUMEN
Induction of back mutations to prototrophy by methylene blue (MB)-sensitized photodynamic (PD) treatment has been studied in wild-type and repair-deficient strains of Salmonella typhimurium carrying either the base-pair substitution mutation hisG46 or the frameshift mutation hisD30529 We found that reversion of the hisG46 mutation was increased in a strain carrying a uvrB deletion and decreased in a strain carrying a recA-type mutation. Reversion of the hisD3052 (frameshift) mutation, on the other hand, was decreased in both uvrB deletion and recA-type strains. The former results are consistent with the hypothesis that the majority of MB-sensitized PD-INDUCED BASE-PAIR SUBSTITUTION MUTATIONS ARIse by a mechanism similar to that currently believed to be involved in UV mutagenesis. The latter results suggest that PD-induced frameshift mutations may arise in some other way, and two possible mechanisms involving sequential action of the excision repair and recombinational repair pathways are considered.
Asunto(s)
Reparación del ADN , Azul de Metileno/farmacología , Mutación , Salmonella typhimurium/efectos de los fármacos , Transcripción Genética , Secuencia de Bases , ADN Bacteriano/biosíntesis , Código Genético , Luz , Genética de Radiación , Recombinación Genética , Salmonella typhimurium/efectos de la radiación , Rayos UltravioletaRESUMEN
The effects of ionizing radiation on cell-cycle progression in lymphoblastoid cell lines derived from ataxia telangiectasia (AT) and Huntington's disease (HD) patients, and from normal individuals, were studied using DNA flow cytometric analysis. A dose of 100 rad gamma irradiation blocked a proportion of normal and HD cells in G1. A higher radiation dose applied to normal cells increased the number of cells blocked in G1 and significantly delayed cells which were in S at the time of irradiation from reaching G2 DNA content. The reduced cumulative mitotic index in irradiated cultures of normal cells 2 h after irradiation suggests that cells in G2 at the time of irradiation are delayed before entering mitosis. After irradiation HD cells responded similarly to normal cells except that a greater proportion of HD cells were blocked in G1. AT cells do not show the normal delay in progression from G1 to S, or from S to G2 in the first cycle after irradiation. The cumulative mitotic index was reduced in irradiated cells, implying that they are delayed in G2. Thus AT cells did not recognize or respond to signals from damaged DNA which in normal and HD cells caused a proportional block in G1 and an S-phase delay. The only point of arrest in cell-cycle progression in irradiated AT cells was in G2.
Asunto(s)
Ataxia Telangiectasia/patología , Ciclo Celular/efectos de los fármacos , Enfermedad de Huntington/patología , Citometría de Flujo , Rayos gamma , Humanos , Interfase/efectos de los fármacosRESUMEN
An approach of general applicability to mammalian radiosensitive mutants has been used in the analysis of gene dosage and complementation in ataxia telangiectasia (A-T). Thymidine residues in DNA of one parental lymphoblastoid cell line were substituted with bromodeoxyuridine before fusion with a second parental cell line, to allow differential staining of the two sets of chromosomes. Following gamma-irradiation, induced chromosome aberrations were scored in diploid and homokaryon cells from each parental line as well as in heterokaryons. Four complementation groups were ascertained among 7 A-T cell lines. Analysis of heterokaryons formed between appropriate combinations of normal, A-T homozygote and A-T heterozygote cells, gave a quantitative measure of gene dosage and demonstrated increasing radiosensitivity with increasing numbers of A-T alleles.
Asunto(s)
Ataxia Telangiectasia/genética , Reparación del ADN , Células Cultivadas , Aberraciones Cromosómicas , Cromosomas/efectos de la radiación , Rayos gamma , Prueba de Complementación Genética , Heterocigoto , Homocigoto , HumanosRESUMEN
Ionizing radiation sensitivity was studied in a series of Huntington's Disease (HD) patients and controls by measurement of radiation-induced chromosome aberrations in lymphocytes and by clonogenic survival of lymphoblastoid cell lines. As a group, HD patients were found to be significantly more radiosensitive than controls (p less than 0.001), but there was an overlap between values for the two groups such that an absolute distinction is not possible. These data are consistent with an association between HD and radiosensitivity but not with identity between HD and a radiosensitive phenotype, so that cellular radiosensitivity cannot be used for individual diagnosis. Analysis of three families including 5 HD patients and 11 first-degree relative confirmed this conclusion and demonstrated that even within a given family presymptomatic diagnosis cannot be based on measurement of radiosensitivity. However, the common association of cellular radiosensitivity with HD probands and their families provides a potential lead to the identification of HD gene(s) and so to an eventual understanding of the aetiopathogenesis of this disease at the molecular level.
Asunto(s)
Genes Dominantes/efectos de la radiación , Enfermedad de Huntington/genética , Línea Celular , Supervivencia Celular/efectos de la radiación , Aberraciones Cromosómicas , Femenino , Humanos , Linfocitos/efectos de la radiación , Masculino , LinajeRESUMEN
Exposure of normal control and ataxia-telangiectasia (A-T) lymphoblastoid cell lines to ionizing radiation gives rise to an increase in the proportion of G2 phase cells. The size and extent of the G2 phase block is greater in A-T cells than in normal cells. Caffeine has a similar overall effect in control and A-T cell lines in reducing the G2 arrest observed after ionizing radiation. While the proportion of cells accumulated in G2 in A-T cells is considerably greater than in controls, addition of caffeine at the time of maximal G2 block brings about a return of G2 phase cell numbers to unirradiated values in 3 hours in both cell types. In normal control cells the caffeine-mediated decrease in G2 cells is reflected by an increase in mitotic cells. These mitotic cells have a higher frequency of chromosome aberrations compared to cells harvested in the absence of caffeine. Similarly in A-T cells addition of caffeine to irradiated cultures, delayed in G2 phase, increased the number of mitotic cells and the frequency of chromosome aberrations.
Asunto(s)
Ataxia Telangiectasia/patología , Cafeína/farmacología , Aberraciones Cromosómicas , ADN/efectos de la radiación , Interfase/efectos de la radiación , Línea Celular , Radioisótopos de Cobalto , Citometría de Flujo , Rayos gamma , Humanos , Técnicas In Vitro , Interfase/efectos de los fármacos , Linfocitos/efectos de la radiación , Factores de TiempoRESUMEN
Five mutants of Salmonella typhimurium strain LT2 trp DI (ColEI)+, initiallly detected because they released little or no colicin when tested on solid medium, proved to be sensitive to ultraviotet light (u.v.). Further testing indicated that one of the mutants was deficient in genetic recombination and was probably a recA-type mutant, while three of the others were deficient in DNA polymerase activity and appeared to be typical polA mutants. The fifth mutant was less sensitive than the others to methyl methanesulphonate, showed reduced proficiency in genetic recombination, and was of approximately normal u.v. mutability. This mutant may be a counterpart of the class known as uvrD in Escherichia coli. All five mutants degraded significantly more of their DNA following exposure to u.v. than did the wild-type strain. The recA-type mutant and the possible uvrD mutant also degraded significantly more of their DNA spontaneously than did the wild-type. Treatment with visible light and acridine orange (photodynamic treatment) cause no significant degradation of DNA in the wild-type strain, a highly significant increase in the extent of DNA degradation in a polA mutant, and a decrease in the extent of degradation in the recA-type mutant.
Asunto(s)
ADN Bacteriano/efectos de la radiación , Salmonella typhimurium/efectos de la radiación , Acridinas , Colicinas/metabolismo , Reparación del ADN , Luz , Mutación , Recombinación Genética , Rayos UltravioletaRESUMEN
To investigate whether predisposition to Wilms' tumour is associated with a particular defect in the handling of DNA damage, cell-lines from families in which the tumour had occurred were tested for sensitivity to a variety of DNA-damaging agents. Lymphoblastoid lines from both Wilms' tumour patients and their first-degree relatives showed increased sensitivity to the cross-linking agent, mitomycin C, but normal sensitivity to ultraviolet (UV) and gamma irradiation. Thus sensitivity to mitomycin C damage can be associated with the Wilms'-tumour-susceptible genotype and could be a genetic factor responsible for the modification of expression of this genotype.
Asunto(s)
Antibióticos Antineoplásicos/farmacología , Neoplasias Renales/genética , Linfocitos/efectos de los fármacos , Mitomicinas/farmacología , Tumor de Wilms/genética , Adulto , Línea Celular , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Niño , Aberraciones Cromosómicas , Susceptibilidad a Enfermedades , Femenino , Rayos gamma , Humanos , Neoplasias Renales/ultraestructura , Linfocitos/efectos de la radiación , Linfocitos/ultraestructura , Masculino , Mitomicina , Intercambio de Cromátides Hermanas/efectos de los fármacos , Intercambio de Cromátides Hermanas/efectos de la radiación , Rayos Ultravioleta , Tumor de Wilms/ultraestructuraRESUMEN
An Australian family is described in which a mild form of xeroderma pigmentosum (XP) is inherited as an autosomal dominant trait. Studies of lymphoblastoid cells and fibroblasts from affected persons demonstrated cellular sensitivity to ultraviolet (UV) light as judged by diminished clonogenicity and higher frequencies of UV induced chromosome aberrations compared to normal controls. After UV irradiation of dominant XP cells, replicative DNA synthesis was depressed to a greater extent than normal and the level of UV induced DNA repair synthesis was lower than that in normal cells. The level of sister chromatid exchanges and the numbers of 6-thioguanine resistant mutants induced by UV irradiation were equal to those found in normal controls. Although two subjects in the family had skin cancers, this dominant form of XP is not apparently associated with high risk, or large numbers, of skin cancers in affected persons.