Brigitte Chapot

Environmental and genetic determinants of aflatoxin-albumin adducts in The Gambia

Aflatoxins together with chronic hepatitis B virus (HBV) infection contribute to the high incidence of hepatocellular carcinoma in developing countries. An understanding of the mechanism of interaction between these factors would provide a strong rationale for developing effective prevention strategies. In this study in The Gambia we examined the effect of environmental (place of residence and timing of sample collection) and host factors (age, sex, HBV status and interindividual variations in carcinogen metabolising enzymes) in determining blood aflatoxin–albumin adduct levels in 357 individuals of whom 181 were chronic HBV carriers. Samples were analysed for aflatoxin‐albumin adducts, HBV status and genotypes of glutathione S‐transferase (GST) M1, GSTT1, GSTP1 and epoxide hydrolase (EPXH). Urine samples were analysed for 6β‐hydroxycortisol:cortisol ratio as a marker of cytochrome P450 (CYP) 3A4 activity. Adduct levels were significantly higher in subjects resident in rural [geometric mean adduct level 34.9 pg aflatoxin B1‐lysine equivalent (28.5–42.8; 95%CI)/mg albumin] than in periurban areas [22.2 pg (14.9–33.4)/mg] and were approximately twice as high in the dry season [mid‐February to March; 83.2 pg (53.3–130.8)/mg] than the wet [July to August; 34.9 pg (28.5‐42.8)/mg]. In contrast, HBV status, CYP3A4 phenotype, GSTT1, GSTP1 and EPXH genotypes were not associated with aflatoxin‐albumin adduct level. However, mean adduct levels were significantly higher in non‐HBV infected subjects with GSTM1 null genotype. The main factors which affect aflatoxin‐albumin adduct levels in this population are environmental, notably place of residence and timing of sample collection. This study further emphasises the priority to reduce aflatoxin exposure in these communities by primary prevention measures. Int. J. Cancer 86:1–7, 2000.

ATM polymorphisms as risk factors for prostate cancer development

The risk of prostate cancer is known to be elevated in carriers of germline mutations in BRCA2, and possibly also in carriers of BRCA1 and CHEK2 mutations. These genes are components of the ATM-dependent DNA damage signalling pathways. To evaluate the hypothesis that variants in ATM itself might be associated with prostate cancer risk, we genotyped five ATM variants in DNA from 637 prostate cancer patients and 445 controls with no family history of cancer. No significant differences in the frequency of the variant alleles at 5557G>A (D1853N), 5558A>T (D1853V), ivs38-8t>c and ivs38-15g>c were found between the cases and controls. The 3161G (P1054R) variant allele was, however, significantly associated with an increased risk of developing prostate cancer (any G vs CC OR 2.13, 95% CI 1.17–3.87, P=0.016). A lymphoblastoid cell line carrying both the 3161G and the 2572C (858L) variant in the homozygote state shows a cell cycle progression profile after exposure to ionising radiation that is significantly different to that seen in cell lines carrying a wild-type ATM gene. These results provide evidence that the presence of common variants in the ATM gene, may confer an altered cellular phenotype, and that the ATM 3161C>G variant might be associated with prostate cancer risk.

The molecular causes of low ATM protein expression in breast carcinoma; promoter methylation and levels of the catalytic subunit of DNA-dependent protein kinase

Aims:  To investigate whether aberrant methylation of the ATM promoter or loss of the catalytic subunit of DNA‐dependent protein kinase (DNA‐PKcs) may be the underlying causes of reduced ATM protein levels often seen in breast tumours.

The methyl methanesulfonate induced S-phase delay in XRCC1-deficient cells requires ATM and ATR

X-ray repair cross-complementing 1 (XRCC1) is required for DNA single-strand break and base excision repair (BER) in human cells. XRCC1-deficient human cells show hypersensitivity to cell killing, increased genetic instability and a significant delay in S-phase progression after exposure to the alkylating agent methyl methanesulfonate (MMS). Using RNAi modulation of XRCC1 levels, we show here that this S-phase delay is associated with significantly increased levels of recombinational repair as visualized by Rad51 focus formation. Using ATM- and ATR-defective cells and an ATM specific kinase inhibitor we demonstrate for the first time that the MMS-induced S-phase checkpoint requires both ATM and ATR. This unique dependency is associated with phosphorylation of ATM/ATR downstream targets or effectors such as SMC1 and Chk1. These results support the hypothesis that after MMS-treatment, the presence of unresolved BER intermediates gives rise to lesions that activate both ATM and ATR and that during the consequent S-phase delay, such intermediates may be repaired by a recombinational pathway which involves the Rad51 protein.

Aflatoxin exposure and cytogenetic alterations in individuals from The Gambia, West Africa

Aflatoxin-albumin adducts in peripheral blood provide a measure of exposure to aflatoxin over the previous 2-3 months. In the present study, the levels of these adducts were determined in a group of individuals from The Gambia, West Africa and were compared in a cross-sectional study to the levels of various cytogenetic alterations (chromosomal aberrations, micronuclei, sister chromatid exchanges) in the same individuals to test whether an increase in genetic damage is associated with an increased exposure in this population. Of 35 subjects tested for aflatoxin-albumin adducts, all but 3 were positive. There were no correlations between the adduct level and the number of cytogenetic abnormalities at the individual level. A comparison of the cytogenetic alterations was made between Gambian individuals and a group of 22 healthy people from Italy where aflatoxin exposure is expected to be low. The levels of structural chromosomal aberrations, sister chromatid exchanges and micronuclei were all higher in the former group. Overall, these data are indicative of a higher exposure to genotoxins in Gambian subjects, one of which are aflatoxins, but suggest that more specific genetic markers of aflatoxin exposure are required to further examine the link between aflatoxin exposure and genetic alterations.

Exposure to hazardous substances in a standard molecular biology laboratory environment: evaluation of exposures in IARC laboratories.

Working in a molecular biology laboratory environment implies regular exposure to a wide range of hazardous substances. Several recent studies have shown that laboratory workers may have an elevated risk of certain cancers. Data on the nature and frequency of exposures in such settings are scanty. The frequency of use of 163 agents by staff working in molecular biology laboratories was evaluated over a period of 4 years by self-administered questionnaire. Of the agents listed, ethanol was used by the largest proportion of staff (70%), followed by ethidium bromide (55%). Individual patterns of use showed three patterns, namely (i) frequent use of a narrow range of products, (ii) occasional use of a wide range of products, and (iii) frequent and occasional use of an intermediate range of products. Among known or suspected carcinogens (International Agency for Research on Cancer Group 1 and 2A, respectively), those most frequently used included formaldehyde (17%), oncogenic viruses (4%), and acrylamide (32%). The type of exposure encountered in research laboratories is extremely diverse. Few carcinogenic agents are used frequently but many laboratory workers may be exposed occasionally to known human carcinogens. In addition, many of the chemicals handled by staff represent a health hazard. The results enabled the staff physician to develop an individual approach to medical surveillance and to draw a personal history of occupational exposures for laboratory staff.