RESUMO
Anti-tumour necrosis factor-α (TNF-α) is used for treatment of severe cases of inflammatory bowel diseases (IBD), including Crohn's disease (CD) and ulcerative colitis (UC). However, one-third of the patients do not respond to the treatment. A recent study indicated that genetically determined high activity of pro-inflammatory cytokines, including interleukin-1ß (IL-1ß), IL-6 and interferon gamma (IFN-γ), are associated with non-response to anti-TNF therapy. Using a candidate gene approach, 21 functional single-nucleotide polymorphisms (SNPs) in 14 genes in the Toll-like receptors, the inflammasome and the IFNG pathways were assessed in 482 and 256 prior anti-TNF naïve Danish patients with CD and UC, respectively. The results were analysed using logistic regression (adjusted for age and gender). Eight functional SNPs were associated with anti-TNF response either among patients with CD (TLR5 (rs5744174) and IFNGR2 (rs8126756)), UC (IL12B (rs3212217), IL18 (rs1946518), IFNGR1 (rs2234711), TBX21 (rs17250932) and JAK2 (rs12343867)) or in the combined cohort of patient with CD and UC (IBD) (NLRP3 (rs10754558), IL12B (rs3212217) and IFNGR1 (rs2234711)) (P<0.05). Only the association with heterozygous genotype of IL12B (rs3212217) (OR: 0.24, 95% CI: 0.11-0.53, P=0.008) among patients with UC withstood Bonferroni correction for multiple testing. In conclusion, Our results suggest that SNPs associated with genetically determined high activity of TLR5 among patients with CD and genetically determined high IL-12 and IL-18 levels among patients with UC were associated with non-response. Further studies will evaluate whether these genes may help stratifying patients according to the expected response to anti-TNF treatment.
Assuntos
Colite Ulcerativa/tratamento farmacológico , Colite Ulcerativa/genética , Doença de Crohn/genética , Interleucina-12/genética , Interleucina-18/genética , Receptor 5 Toll-Like/genética , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Estudos de Coortes , Feminino , Humanos , Doenças Inflamatórias Intestinais/tratamento farmacológico , Doenças Inflamatórias Intestinais/genética , Interferon gama/genética , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único/genética , Adulto JovemRESUMO
Antitumor necrosis factor-α (TNF-α) is used for treatment of severe cases of inflammatory bowel diseases (IBD), including Crohn's disease (CD) and ulcerative colitis (UC). However, one-third of the patients do not respond to the treatment. Genetic markers may predict individual response to anti-TNF therapy. Using a candidate gene approach, 39 mainly functional single nucleotide polymorphisms (SNPs) in 26 genes regulating inflammation were assessed in 738 prior anti-TNF-naive Danish patients with IBD. The results were analyzed using logistic regression (crude and adjusted for age, gender and smoking status). Nineteen functional polymorphisms that alter the NFκB-mediated inflammatory response (TLR2 (rs3804099, rs11938228, rs1816702, rs4696480), TLR4 (rs5030728, rs1554973), TLR9 (rs187084, rs352139), LY96 (MD-2) (rs11465996), CD14 (rs2569190), MAP3K14 (NIK) (rs7222094)), TNF-α signaling (TNFA (TNF-α) (rs361525), TNFRSF1A (TNFR1) (rs4149570), TNFAIP3(A20) (rs6927172)) and other cytokines regulated by NFκB (IL1B (rs4848306), IL1RN (rs4251961), IL6 (rs10499563), IL17A (rs2275913), IFNG (rs2430561)) were associated with response to anti-TNF therapy among patients with CD, UC or both CD and UC (P ⩽ 0.05). In conclusion, the results suggest that polymorphisms in genes involved in activating NFκB through the Toll-like receptor (TLR) pathways, genes regulating TNF-α signaling and cytokines regulated by NFκB are important predictors for the response to anti-TNF therapy among patients with IBD. Genetically strong TNF-mediated inflammatory response was associated with beneficial response. In addition, the cytokines IL-1ß, IL-6 and IFN-γ may be potential targets for treating patients with IBD who do not respond to anti-TNF therapy. These findings should be examined in independent cohorts before these results are applied in a clinical setting.
Assuntos
Doenças Inflamatórias Intestinais/tratamento farmacológico , Doenças Inflamatórias Intestinais/genética , NF-kappa B/metabolismo , Polimorfismo de Nucleotídeo Único/genética , Transdução de Sinais/genética , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Estudos de Coortes , Dinamarca , Feminino , Humanos , Doenças Inflamatórias Intestinais/metabolismo , Masculino , Pessoa de Meia-Idade , NF-kappa B/antagonistas & inibidores , Polimorfismo de Nucleotídeo Único/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismo , Adulto JovemRESUMO
Repeated percutaneous applications of 7,12-dimethylbenz(a)anthracene on weaning DBA/2 and ST/a mice induced 100% leukemias with short latency periods. Endogenous C-type viruses were activated during the treatment as evidenced by (a) increased expression of the murine leukemia virus major core protein, p30, in blood and spleens and (b) increased frequency of detection of ecotropic virus by cocultivation of the splenocytes with SC-1 cells. The treatment did not affect p30 expression in several nonlymphoid organs, and detection of xenotropic viruses in the splenocytes was decreased. Virus expression did not correlate with the progression of disease in that (a) high p30 levels were generally found in mice with relatively low spleen weights and (b) p30 levels had no obvious connection to survival of the individual. 7,12-Dimethylbenz(a)anthracene treatment had little influence on p30 expression in spleens and blood from C3H and BALB/c mice, which are less sensitive to 7,12-dimethylbenz(a)anthracene-induced leukemogenesis. The results indicate an association of C-type virus activation with chemical induction of leukemia but do not necessarily imply an etiological role of the virus in the disease.
Assuntos
9,10-Dimetil-1,2-benzantraceno/farmacologia , Benzo(a)Antracenos/farmacologia , Leucemia Experimental/etiologia , Retroviridae/efeitos dos fármacos , Animais , Coração/microbiologia , Leucemia Experimental/sangue , Leucemia Experimental/microbiologia , Masculino , Camundongos , Camundongos Endogâmicos , Retroviridae/isolamento & purificação , Especificidade da Espécie , Baço/microbiologia , Proteínas Virais/sangue , Replicação Viral/efeitos dos fármacosRESUMO
The content of ribo- and deoxyribonucleotide triphosphates has been measured in asynchronous and synchronized populations of Tetrahymena pyriformis. The concentration of ribonucleotide triphosphates does not vary through the cell cycle. It seems to be a rule that the ATP pool is reduced during division only in cell types which also reduce macromolecular syntheses and respiration. The content of deoxyribonucleotide triphosphates varies through the cell cycle. The pools are maximal during the S-period, but measurable amounts are present in all phases.
Assuntos
Desoxirribonucleotídeos/metabolismo , Ribonucleotídeos/metabolismo , Tetrahymena pyriformis/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Divisão Celular , Cromatografia em Camada Fina , DNA/biossíntese , Mitose , Espectrofotometria Ultravioleta , Timidina/metabolismo , Fatores de TempoRESUMO
We have explored the structural features of cobalamin binding proteins by peptide mapping. The present report is a comparison of the radioiodinated tryptic peptides of intrinsic factor, transcobalamin and haptocorrin from man, hog and rabbit. The results show that the homology between analogous proteins from different species is close for intrinsic factor and transcobalamin and weaker for haptocorrin. The results also suggest the existence of one or more regions which, with minor changes, are conserved among all proteins investigated. This implies a common evolutionary origin for all the cobalamin binding proteins studied.
Assuntos
Proteínas Sanguíneas/análise , Fator Intrínseco/análise , Fragmentos de Peptídeos/isolamento & purificação , Transcobalaminas/análise , Animais , Humanos , Radioisótopos do Iodo , Desnaturação Proteica , Coelhos , Especificidade da Espécie , Suínos , TripsinaRESUMO
The XPD protein has a dual function, both in nucleotide excision repair and in basal transcription. We have studied the role of two nucleotide substitutions in the XPD gene, one in exon 23 leading to an amino acid substitution (Lys751Gln) and one silent in exon 6 in relation to basal cell carcinoma (BCC). Both are two-allele polymorphisms, with the nucleobases A and C at the given positions. We genotyped psoriasis patients with and without BCC and nonpsoriatic persons with and without BCC (4 x 20 persons). The choice to study psoriasis patients was motivated by their high genotoxic exposure via treatment and their high relative rate of early BCC. Subjects carrying two A alleles (AA genotype) in exon 23 were at 4.3-fold higher risk of BCC than subjects with two C alleles (95% CI, 0.79-23.57). In addition, the mean age at first skin tumor for BCC cases with the AA genotype was significantly lower than the mean age for BCC cases with the AC or CC genotype (P = 0.012). Thus, the variant C-allele of exon 23 may be protective. The exon 6 genotype was associated with the risk of BCC among the psoriasis patients; psoriatics carrying two A alleles in exon 6 were at 5.3-fold higher risk of BCC than psoriatics with two C alleles (95% CI, 0.78-36.31). For the psoriatics, the mean age at onset of BCC for cases with the AA genotype was marginally lower than the mean age for cases with genotype AC or CC (P = 0.060). Our results raise the possibility that the polymorphisms in the XPD gene may be contributing factors in the risk of BCC development. They are, therefore, important candidates for future studies in susceptibility to cancer.
Assuntos
Idade de Início , Carcinoma Basocelular/genética , DNA Helicases/genética , Reparo do DNA/genética , Proteínas de Ligação a DNA , Polimorfismo Genético/genética , Proteínas/genética , Fatores de Transcrição , Adenina , Adulto , Fatores Etários , Alelos , Intervalos de Confiança , Citosina , Éxons/genética , Feminino , Predisposição Genética para Doença , Variação Genética/genética , Genótipo , Glutamina/genética , Humanos , Lisina/genética , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Psoríase/genética , Fatores de Risco , Transcrição Gênica/genética , Proteína Grupo D do Xeroderma PigmentosoRESUMO
We have investigated the formation of strand-breaks following UVC irradiation in lymphocytes from psoriasis patients with or without basal cell carcinoma (BCC). Isolated lymphocytes were irradiated with UVC light at a dose of 3.6 J/m(2), and the level of DNA strand-breaks were measured 25 min after the irradiation by the alkaline comet assay. The generation of strand-breaks following UVC irradiation indicates DNA-repair-mediated incisions, as UVC light does not generate strand-breaks per se. We found that psoriasis patients with BCC had more DNA-repair incisions than non-cancer patients. The incision level correlated to two polymorphisms of the XPD gene. At present, it is not clear if the association is a primary effect that is related to differences of the XPD protein. Genes encoding for other repair proteins, namely XRCC1, ERCC1, and LIG1 are located close to the XPD gene, and it is possible that the association is due to a cosegregation with a polymorphism in one of these genes.
Assuntos
Carcinoma Basocelular/complicações , Carcinoma Basocelular/genética , Dano ao DNA/efeitos da radiação , DNA Helicases , Reparo do DNA/genética , Proteínas de Ligação a DNA , Linfócitos/efeitos da radiação , Psoríase/complicações , Psoríase/genética , Neoplasias Cutâneas/genética , Fatores de Transcrição , Alelos , Carcinoma Basocelular/patologia , Células Cultivadas , Ensaio Cometa , Dano ao DNA/genética , Reparo do DNA/efeitos da radiação , Relação Dose-Resposta à Radiação , Éxons/genética , Feminino , Ligação Genética/genética , Predisposição Genética para Doença/genética , Humanos , Linfócitos/metabolismo , Linfócitos/patologia , Masculino , Polimorfismo Genético/genética , Proteínas/genética , Psoríase/patologia , Neoplasias Cutâneas/complicações , Neoplasias Cutâneas/patologia , Raios Ultravioleta , Proteína Grupo D do Xeroderma PigmentosoRESUMO
We have previously shown that high DNA repair capacity protects psoriasis patients against chemically induced basal cell carcinoma [Dybdahl et al. Mutat. Res. 433 (1999) 15-22]. We have used the same study persons to investigate the correlation between expression of eight genes involved in nucleotide excision repair and DNA repair capacity. mRNA levels of XPA, XPB, XPC, XPD, XPF, XPG, CSB and ERCC1 in primary lymphocytes from 33 individuals were quantified by dot-blots and normalized to beta-actin. ERCC1 and XPD mRNA quantities were highly correlated (r=0.89; P<10(-11)) while XPA, XPB, XPC, XPG, XPFand CSB mRNAs were moderately correlated (r=0.2-0.7). Thus, the mRNA expressions seem to fall in at least two groups. There was a three to sevenfold variation in the expression levels of the mRNAs. This is in contrast to the more than a hundredfold variation in mRNA levels reported in cancer patients.DNA repair capacity was measured in a host cell reactivation assay, where primary lymphocytes were transfected with an UV-irradiated plasmid encoding firefly-luciferase. Only ERCC1 and XPD mRNA levels correlated with the DNA repair capacity (P<0.03). In order to see if ERCC1 or XPD activity was limiting for DNA repair, we cotransfected with plasmids encoding NER genes, thus over-expressing either XPB, XPC, XPD, CSB or ERCC1 in the host cell reactivation assay. Only XPB over-expression increased DNA repair capacity. Thus, there is no indication that neither XPD nor ERCC1 limits the DNA repair capacity. However, our results indicate that ERCC1 and XPD mRNA levels may be used as a proxy for DNA repair capacity in lymphocytes.
Assuntos
Reparo do DNA/genética , Linfócitos/metabolismo , RNA Mensageiro/metabolismo , Sequência de Bases , Linhagem Celular Transformada , Primers do DNA , Humanos , RNA Mensageiro/genética , TransfecçãoRESUMO
We have studied DNA repair in patients with psoriasis aiming at investigating the importance of repair in chemically induced cancer. An increased risk of non-melanoma skin cancer has been observed in psoriasis patients extensively treated with tar, methotrexate and photochemotherapy (psoralen + UVA). We measured the DNA repair capacity (DRC) by a host cell reactivation (HCR) assay in lymphocytes from psoriasis patients with and without basal cell cancer and non-psoriatic persons with and without basal cell cancer (4 x 20 study persons). Among psoriasis patients we observed a significant lower DRC in patients with skin cancer compared to patients without skin cancer (P = 0.015; Mann-Whitney, one-sided). Using the median of the healthy control group (group 4) as a cutoff value to divide the psoriasis patients into groups of high and low repair, we found that individuals who had a low repair capacity had a 6.4-fold increased skin cancer risk compared to individuals with high repair (95% confidence interval (CI), 1.44-28.5). The level of DNA repair was correlated with the age at which the psoriasis patients got their first skin cancer. The lower the level of DNA repair, the earlier the psoriasis patients had their first skin tumor (P = 0.070 Spearman; one-sided). Psoriasis patients without BCC had marginally higher repair than healthy controls (P = 0.11, Mann-Whitney, two-sided). We found no difference between BCC patients without psoriasis and healthy controls. In conclusion, these findings suggest a protective role of DNA repair in a predominantly chemically induced cancer.
Assuntos
Carcinoma Basocelular/genética , Reparo do DNA/genética , Psoríase/complicações , Neoplasias Cutâneas/genética , Adulto , Fatores Etários , Carcinoma Basocelular/induzido quimicamente , Carcinoma Basocelular/complicações , DNA de Neoplasias/efeitos da radiação , Dinamarca , Feminino , Ficusina/farmacologia , Humanos , Linfócitos/metabolismo , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Neoplasias Cutâneas/induzido quimicamente , Estatísticas não Paramétricas , Transfecção/genética , Células Tumorais Cultivadas , Terapia UltravioletaRESUMO
Quadruples of skin cancer patients with and without psoriasis and referents with and without psoriasis (4 x 20 study persons) were identified and examined for DNA damage by single cell gel electrophoresis (comet-assay) and DNA-repair by UV-induced unscheduled DNA synthesis (UDS) in mononuclear blood cells (lymphocytes and monocytes). DNA damage (strand breaks and alkaline labile sites) as assessed by the comet assay and DNA repair as assessed by UDS were significantly associated with the season in which blood sampling took place. This variation might be explained by an increased exposure to solar radiation. When the comet tail moment data were stratified by sampling period, an interaction between psoriasis and skin cancer was detected, with patients with psoriasis and skin cancer exhibiting more DNA damage. Patients with psoriasis and skin cancer also had lower UDS compared to healthy study persons, suggesting that the more DNA damage may be caused by a lower rate of DNA repair. In all study persons, the extent of UDS correlated positively with the amount of DNA damage determined by the comet assay.
Assuntos
Carcinoma Basocelular/genética , Dano ao DNA , Reparo do DNA , Psoríase/complicações , Estações do Ano , Neoplasias Cutâneas/genética , Contagem de Células Sanguíneas , Carcinoma Basocelular/complicações , Dano ao DNA/efeitos da radiação , Reparo do DNA/efeitos da radiação , Feminino , Humanos , Linfócitos/metabolismo , Masculino , Pessoa de Meia-Idade , Monócitos/metabolismo , Neoplasias Cutâneas/complicaçõesRESUMO
Three major pathways, nucleotide excision repair (NER), base excision repair (BER) and O6-methylguanine-DNA methyltransferase (MGMT), are responsible for the removal of most adducts to DNA and thus for the survival of cells influenced by deoxyribonucleic acid (DNA) adduct-forming chemicals. We have evaluated host cell reactivation and cell survival of wild type Chinese hamster ovary cells and of mutants in the NER-genes ERCC1, ERCC2, and ERCC4 after treatment with the methylating compounds dimethylsulfate and methylnitrosourea. No effect of the three genes could be demonstrated, i.e., survival and host cell reactivation after methylation damage in the mutants and the wild type cells were similar. Gene-specific repair experiments confirmed the proficient removal of methyl lesions. We conclude that the three nucleotide excision repair genes are immaterial to the repair of methylation damage. This suggests that NER does not play a role in the removal of methylation in mammalian cells and that BER and MGMT are responsible for the survival of such cells, when they are challenged with methylation of DNA.
Assuntos
Reparo do DNA , Proteínas de Ligação a DNA/genética , Endonucleases , Mutação , Proteínas/genética , Animais , Células CHO , Cricetinae , Adutos de DNA/genética , Dano ao DNA , Metilação de DNA , FemininoRESUMO
OBJECTIVES: The purpose of this study was to compare different methods for calculating maximal allowable concentrations of potentially carcinogenic substances in indoor air. Benzene, tetrachloroethylene, trichloroethylene, and vinyl chloride were selected as the model substances. METHODS: Estimates were used of carcinogenic potency from quantitative risk assessment, lowest observable effect levels (LOEL) from animal experiments and epidemiologic studies combined with safety factors, and estimation from occupational exposure limits with safety factors. The estimates were compared with actual concentrations in buildings in Denmark. RESULTS: Concentrations of benzene, tetrachloroethylene, trichloroethylene, and vinyl chloride of the order of 10, 20, 200, and 40 ppb, respectively, in indoor air were found to correspond to a 10(-4) lifetime risk of cancer. CONCLUSIONS: The estimated maximal allowable concentrations of carcinogenic compounds obtained from indoor air by quantitative risk assessment using a lifetime risk of 10(-4) or using LOEL values and suitable safety factors appear to be comparable and reasonable. Calculations based on occupational exposure limits and safety factors generally give comparable or somewhat higher values. Using a lifetime risk of 10(-6) for quantitative risk assessment does not seem reasonable considering the risks associated with the activities of everyday life.
Assuntos
Poluição do Ar em Ambientes Fechados/análise , Carcinógenos/análise , Animais , Dinamarca , Humanos , Concentração Máxima Permitida , Exposição Ocupacional/análise , Fatores de RiscoRESUMO
The major core protein, p30, of mouse C-type viruses was quantitated radioimmunologically in lymphoid organs and blood from inbred strains of mice. The concentration of p30 in thymus and spleen had a weak and moderate correlation, respectively, to leukemia frequency. In contrast, the concentration of p30 in blood from mice with a high incidence of leukemia (strains AKR and C58) was 100-fold increased at 2 months of age compared with 10 strains with a low incidence of the disease. The SJL mice, which have a high incidence of reticulum cell neoplasms, showed generally elevated, but variable, values. The high concentration in AKR blood developed during the first weeks of life. Approximately one-third of the DBA/2 mice had elevated levels after 4 to 5 months, whereas the values from mice of the 129 strain were low irrespective in their age. The major part of p30 appeared to be associated with the erythrocytes. The concentration of p30 in the blood seems to reflect the presence of replicating virus in mice. It identifies among the inbred strains a high leukemia group one-half year prior to disease.
Assuntos
Leucemia/epidemiologia , Camundongos Endogâmicos/sangue , Retroviridae , Proteínas Virais/sangue , Fatores Etários , Animais , Eritrócitos/análise , Camundongos , Camundongos Endogâmicos AKR , Camundongos Endogâmicos DBA , Especificidade da Espécie , Baço/análise , Timo/análise , Proteínas Virais/análiseRESUMO
Ecotropic C-type retroviruses isolated from both normal and dimethylbenzanthracene-treated DBA/2 mice could be classified into three major groups, Ea, Eb, and Ec, that differed in structure and biology. Weanling DBA/2 mice were generally free of viruses, but a fraction of adult individuals became virus positive and were apparently selectively associated with a high expression of the Eb viruses. Some of the ecotropic viruses from DBA/2 mice acted as exogenous pathogens. They caused viremia and a moderate incidence of leukemia when injected into C3H and ST/a mice. In addition, they caused an appreciable number of early deaths without signs of malignancy. To evaluate the natural role of the viruses, we studied the survival of spontaneously viremic and nonviremic DBA/2 mice. The viremic animals as a group were characterized by a significantly reduced life-span that was not related to neoplasia. These observations indicated that endogenous C-type retroviruses can be pathogenic without preselection of the host for disease. They also emphasize that endogenous viruses, like their exogenous counterparts, can have a broader pathogenic spectrum than normally appreciated.
Assuntos
Camundongos Endogâmicos DBA/microbiologia , Retroviridae/patogenicidade , Infecções Tumorais por Vírus/microbiologia , 9,10-Dimetil-1,2-benzantraceno , Animais , Leucemia Experimental/microbiologia , Camundongos , Camundongos Endogâmicos/microbiologia , Retroviridae/genética , Retroviridae/isolamento & purificação , Especificidade da EspécieRESUMO
We have studied the entry of murine retrovirus into mouse fibroblasts by following the fate of both radioactively (protein) labeled virus particles and infectious virus particles. Physical and infectious particles bound to the cell surface with a half time of 1.5-2 hr. Both types of particles were internalized with a half time of approximately 3 hr as measured by the resistance to externally added proteases. The binding proceeded both at 37 and 0 degrees, whereas the internalization was blocked at 0 degrees. The internalized physical particles followed two routes: they either were degraded or remained stable in the cell. Degradation was blocked by lysosomotropic bases and is therefore believed to occur in the lysosomes. Infection could also be inhibited by lysosomotropic bases when present in the first hours after the internalization, indicating that the infectious route also is leading through the lysosomes or another acidic compartment of the cell.
Assuntos
Fibroblastos/microbiologia , Retroviridae/fisiologia , Animais , Linhagem Celular , Membrana Celular/microbiologia , Cinética , Lisossomos/metabolismo , Lisossomos/microbiologia , Camundongos , Retroviridae/metabolismo , Temperatura , Tripsina/farmacologia , Proteínas Virais/metabolismoRESUMO
Ecotropic murine leukemia viruses isolated from normal and carcinogen-treated DBA/2 mice can be classified into three main groups that differ in structure and biology. Two groups, called Ea and Eb, consist of N-tropic viruses related to the standard endogenous ecotropic virus of AKR mice. Ea viruses replicate with reduced efficiency in cell lines derived from C3H mice, while Eb viruses essentially replicate normally in these cells. As elsewhere reported, Ea viruses appear apathogenic in C3H mice, while Eb viruses cause a moderate incidence of late leukemias. The biological differences are associated with modulations of the fine structure of the gag gene-encoded proteins. A third group of viruses, called Ec, is clearly more diverged. They differ extensively from Ea and Eb viruses in the products of the gag and env gene, and are related to Rauscher leukemia virus. Ec viruses are NB-ecotropic; they replicate efficiently in all mouse cells tested, and induce leukemias in C3H mice with shorter latency periods than Eb viruses. Since published nucleic acid hybridization data indicate that DBA/2 mice only carry one ecotropic provirus, we assume that the DBA/2 viruses represent a developmental series of variants evolving during the life of the animals.
Assuntos
Retroviridae/isolamento & purificação , Proteínas Virais/análise , Animais , Suscetibilidade a Doenças , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Camundongos Endogâmicos DBA , Peso Molecular , Retroviridae/fisiologia , Especificidade da Espécie , Proteínas do Envelope Viral , Replicação ViralRESUMO
The XPD gene product has a dual function in basal transcription and in nucleotide excision repair. We have previously reported that two polymorphisms in the gene, one silent mutation in codon 156 of exon 6 and one giving rise to a Lys-->Gln substitution in codon 751 of exon 23, showed signs of being associated with basal cell carcinoma in a Scandinavian study group of psoriasis patients and non-psoriatics with and without basal cell carcinoma [Dybdahl, Vogel, Frentz, Wallin and Nexø (1999) Cancer Epidemiol. Biomark. Prev., 8, 77-81]. In both polymorphisms, the CC genotype appeared to be protective against basal cell carcinoma. Here, we have genotyped an American study group of basal cell carcinoma patients and controls without skin cancer for the two polymorphisms. In addition, we studied an A-->G polymorphism in codon 312 of exon 10, which results in an Asp-->Asn substitution in a conserved region of XPD. In the whole study group, subjects carrying the AA and AC genotype in exon 6 were at 1.9-fold higher risk of basal cell carcinoma (P = 0.062, CI 0.96-3.75). If only subjects without a family history of non-melanoma skin cancer were included, subjects carrying AA or AC genotype were at 3.3-fold higher risk of basal cell carcinoma (P = 0.007, CI 1.35-8.18). Among subjects with a family history of non-melanoma skin cancer, subjects with an AG or AA genotype in codon 312 of exon 10 were at 5.25-fold increased risk of basal cell carcinoma (P = 0.027, CI 1.15-23.93). A protective effect of the CC genotype in exon 23 could not be confirmed. Cases with a family history of skin cancer had statistically significantly different allele frequencies of the polymorphisms in exon 6 and exon 10 from cases without family history of non-melanoma skin cancer. Our results indicate that the exon 6(A) allele is a risk factor in basal cell carcinoma.
Assuntos
Carcinoma Basocelular/genética , DNA Helicases , Reparo do DNA/genética , Proteínas de Ligação a DNA , Polimorfismo Genético , Proteínas/genética , Neoplasias Cutâneas/genética , Fatores de Transcrição , Adulto , Alelos , Estudos de Casos e Controles , Códon , Éxons , Feminino , Predisposição Genética para Doença , Humanos , Masculino , Pessoa de Meia-Idade , Proteína Grupo D do Xeroderma PigmentosoRESUMO
Somatic cell gene mutation arising in vivo may be considered to be a biomarker for genotoxicity. Assays detecting mutations of the haemoglobin and glycophorin A genes in red blood cells and of the hypoxanthine-guanine phosphoribosyltransferase and human leucocyte antigenes in T-lymphocytes are available in humans. This MiniReview describes these assays and their application to studies of individuals exposed to genotoxic agents. Moreover, with the implementation of techniques of molecular biology mutation spectra can now be defined in addition to the quantitation of in vivo mutant frequencies. We describe current screening methods for unknown mutations, including the denaturing gradient gel electrophoresis, single strand conformation polymorphism analysis, heteroduplex analysis, chemical modification techniques and enzymatic cleavage methods. The advantage of mutation detection as a biomarker is that it integrates exposure and sensitivity in one measurement. With the analysis of mutation spectra it may thus be possible to identify the causative genotoxic agent.
Assuntos
Monitoramento Ambiental , Mutação , Humanos , Mutagênese/fisiologiaRESUMO
The cytotoxic and mutagenic effect of the bifunctional alkylating agent nitrogen mustard (HN2) was examined. Primary human lymphocytes were exposed to graded doses of HN2 in vitro and relative survival was determined. Mutation induction at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus was measured by cloning the exposed T-cells in microtitre plates in the presence and absence of 6-thioguanine (TG). The IC50-value determined for 30 min exposure to HN2 was 1.34 microM. The mutant frequencies (MF) in exposed T-cell cultures were 10-fold (2 microM HN2) to 32-fold (4 microM HN2) higher than those of unexposed cultures (median values). Nitrogen mustard-mediated mutagenesis is discussed in terms of the current ideas about DNA damage and repair.