Arsenic-related DNA copy-number alterations in lung squamous cell carcinomas.

BACKGROUND: Lung squamous cell carcinomas (SqCCs) occur at higher rates following arsenic exposure. Somatic DNA copy-number alterations (CNAs) are understood to be critical drivers in several tumour types. We have assembled a rare panel of lung tumours from a population with chronic arsenic exposure, including SqCC tumours from patients with no smoking history. METHODS: Fifty-two lung SqCCs were analysed by whole-genome tiling-set array comparative genomic hybridisation. Twenty-two were derived from arsenic-exposed patients from Northern Chile (10 never smokers and 12 smokers). Thirty additional cases were obtained for comparison from North American smokers without arsenic exposure. Twenty-two blood samples from healthy individuals from Northern Chile were examined to identify germline DNA copy-number variations (CNVs) that could be excluded from analysis. RESULTS: We identified multiple CNAs associated with arsenic exposure. These alterations were not attributable to either smoking status or CNVs. DNA losses at chromosomes 1q21.1, 7p22.3, 9q12, and 19q13.31 represented the most recurrent events. An arsenic-associated gain at 19q13.33 contains genes previously identified as oncogene candidates. CONCLUSIONS: Our results provide a comprehensive approach to molecular characteristics of the arsenic-exposed lung cancer genome and the non-smoking lung SqCC genome. The distinct and recurrent arsenic-related alterations suggest that this group of tumours may be considered as a separate disease subclass.

Smoking habit and genetic factors associated with lung cancer in a population highly exposed to arsenic.

In order to find some relationship between genetic differences in metabolic activation and detoxification of environmental carcinogens and host susceptibility to chemically induced cancers, we have investigated the distribution of the GSTM1 null genotype and CYP450 *1A1 MspI polymorphism in lung cancer patients and healthy volunteers of the second region in the north of Chile highly exposed to arsenic. The main sources of environmental arsenic exposure in Chile are copper smelting and drinking water, specially in the second region, the most important copper mining region in the world that shows the highest lung cancer mortality rate in the country (35/100.00). The population of Antofagasta, the main city of the region was exposed between 1958 and 1970 to arsenic concentrations in drinking water of 860 microg/m3, presently declining to 40 microg/m3. For men the MspI CYP1A1 *2A genotype was associated with a highly significant estimated relative lung cancer risk (O.R. = 2.60), but not GSTM1 by itself. The relative lung cancer risk for the combined 2A/null GSTM1 genotypes was 2.51, which increased with the smoking habits (O.R. = 2.98). In the second region the cancer mortality rate for As associated cancers, might be related at least part to differences in As biotransformation. In this work we demonstrate that genetic biomarkers such as CYP1A1 2A and GSTM1 polymorphisms in addition to DR70 as screening biomarkers might provide relevant information to identify individuals with higher risk for lung cancer, due to arsenic exposure.

CYP1A1 and GSTM1 genetic polymorphisms in lung cancer populations exposed to arsenic in drinking water.

Region II of Chile is the most important copper mining area in the world and it shows the highest lung cancer mortality rate in the country (35/100,000). The population in Antofagasta, the main city of Region II, was exposed from 1958 to 1970 to 860 microg m(-3) arsenic (As) in drinking water and has currently been declining to 40 microg m(-3). Glutathione serves as a reducing agent and glutathione S-transferase (GST) may have an important role in As methylation capacity and body retention. In the current study, the null genotype of GSTM1 and the MspI polymorphism of CYP450 1A1 were investigated in lung cancer patients and in healthy volunteers of Region II. In males, the 2A genotype of MspI represented a highly significant estimated relative lung cancer risk (OR=2.60). Relative lung cancer risk for the combined 2A/null GSTM1 genotypes was 2.51, which increased with the smoking habit (OR=2.98). In Region II, the cancer mortality rate for As-associated cancers at least partly might be related to differences in As biotransformation. Genetic biomarkers such as 2A and GSTM1 polymorphisms in addition to DR70 as screening biomarkers might provide relevant information to identify individuals with a high risk for lung cancer as prevention and protection actions to protect public health.

Susceptibility and exposure biomarkers in people exposed to PAHs from diesel exhaust.

Xenobiotic metabolizing enzymes, especially CYP1A1 and GSTM1, are involved in the activation and conjugation of PAHs and are controlled by polymorphic genes. PAHs released from diesel emissions in many cities of the world, especially in developing countries, contribute significantly to the toxic effects of airborne inhalable particles. We have evaluated the gene-environment interaction in Santiago of Chile, studying the contribution of CYP1A1 and GSTM1 polymorphisms on 1-OH-P urinary levels used as the PAHs exposure biomarker. The study was performed on 59 diesel exposed (38 diesel revision workers and 21 subjects working in an urban area as established street vendors) and 44 non-exposed subjects living in a rural area. The 1-OH-P urinary levels of the urban (P=0.043) and rural (P=0.040) populations showed, without considering the genotypes, significant differences between smokers and non-smokers, but no significant differences were found between smokers and non-smokers among the diesel plant workers (P=0.33). Non-smoking subjects of the diesel plants and the urban area showed similar 1-OHP levels (P=0.466) which were significantly higher than those of the subjects living in the rural area (P<0.05). When 1-OH-P levels were related with genotypes, an association was observed for the CYP1A1*2A genotype, so that the diesel-exposed workers carrying the CYP1A1*2A allele showed significantly higher 1-OH-P levels than the subjects from the rural area with the same genotype (P=0.008). On the other hand, there was no significant correlation between urinary 1-OH-P levels and GSTM1 null genotype, although higher levels of the urinary metabolite were found in individuals carrying the combined CYP1A1*2A and GSTM1 null genotype (P=0.055). These results may suggest an association between levels of the exposure biomarker 1-OH-P and presence of the CYP1A1*2A genotype, a potential genetic susceptibility biomarker which might be useful in identifying individuals at higher risk among people exposed to high PAH levels in diesel exhaust.

[Indoor air pollution in a zone of extreme poverty of La Pintana, Santiago-Chile]. / Contaminación intradomiciliaria en un sector de extrema pobreza de la comuna de la pintana.

BACKGROUND: Indoor pollution can be an important risk factor for human health, considering that people spend more than 60% of their time in their houses. AIM: To investigate indoor pollution in a zone of extreme poverty in Metropolitan Santiago. MATERIAL AND METHODS: During 24 h, carbon monoxide (CO), sulfur dioxide (SO2), respirable particulate matter (PM10), polycyclic aromatic hydrocarbons absorbed in PM5, temperature and humidity, were measured in the interior of 24 houses in a La Pintana, Santiago. RESULTS: The higher pollutant concentrations were observed during hours when heating was used, in houses that used coal (mean PM10 250 micrograms/m3, CO 42 ppm, SO2 192 pph) or firewood (mean PM10 489 micrograms/m3, CO 57 ppm, SO2 295 pph). In all houses, polycyclic aromatic hydrocarbons were detected and they came from the interior of the house and not from external filtered air. Coal, firewood and cigarette smoke were important sources of carcinogenic and kerosene and gas were sources of non carcinogenic polycyclic aromatic hydrocarbons. CONCLUSIONS: In the houses studied, the population was exposed to an accumulation of highly toxic pollutants, caused by a lack of ventilation. A high relative humidity also contributed to the growth of biological pollutants.

Polycyclic aromatic hydrocarbon levels and mutagenicity of inhalable particulate matter in Santiago, Chile.

The air in Santiago, Chile, is among the most highly polluted in the world. Due to the high levels of pollutants and the high incidence of respiratory diseases, especially in the most susceptible groups, Santiago has been declared a saturated zone for PM(10), O(3), and CO. The aim of this work was to investigate polycyclic aromatic hydrocarbon levels and mutagenic activity of Santiago s fine and coarse fractions of inhalable particles. The levels of 16 polycyclic aromatic hydrocarbons (PAHs) were determined by high-performance liquid chromatography in organic extracts from respirable particles (OERP). Respirable particulate matter (fine and coarse) contains high levels of PAHs including six mutagenic ones classified by the IARC as carcinogenic, which represented at least 45% of the total PAH concentration. A seasonal effect was observed, with higher values in months with lower temperatures. Although a significant decline of PAH levels in OERP was observed in the last years, the levels of carcinogenic PAHs are still higher than those reported in cities of the United States, Australia, and Europe. OERP were highly mutagenic and contained direct and indirect mutagens, which produced both frameshift and base substitution mutations in Salmonella typhimurium. In addition, organic extracts from total suspended particles were also highly mutagenic at the tk locus in h1A1v2 human lymphoblasts in culture. In spite of the important decrease in PAHs in the period 1991-1996, direct mutagenic response has not changed significantly, suggesting that the levels of direct mutagenic pollutants (e.g., nitroarenes) have not decreased considerably during the last years. These results suggest a risk for Santiago s inhabitants since pollutants adsorbed in inhalable particles are highly mutagenic and can damage DNA.

Trends of polycyclic aromatic hydrocarbon levels and mutagenicity in Santiago's inhalable airborne particles in the period 1992-1996.

Trends of polycyclic aromatic hydrocarbons (PAHs) for 1992-1996 (cold season) and their mutagenic activity were investigated in organic extracts from the Santiago, Chile, inhalable particles (PM(10)). The highest PAH concentrations were observed in 1992 and declined dramatically in the following years. During this period, total PAHs decreased 85%, carcinogenic PAHs 82%, and benzo[a]pyrene, the most potent carcinogen, 85%. In spite of this significant decrease, PAH levels in respirable particles were higher than those reported in recent studies in Australia, Europe, and the United States. PAH profiles were analyzed by principal component (PC) analysis and Pearson correlation analysis. PC1 represents 71% of the variance, suggesting that most PAHs might originate predominantly from one main generic source. Higher correlations were obtained for the major carcinogenic PAHs. Most of the samples assayed were highly mutagenic to Salmonella typhimurium both in the presence and in the absence of metabolic activation system (S9), especially in the coarse fraction, but direct mutagenicity did not decline significantly. Incubation of calf thymus DNA with organic extracts from particulate matter and xanthine oxidase allowed the detection of five nitro-PAH-DNA adducts. Thus, nitroarenes might play an important role in the mutagenic activity of inhalable particles in Santiago, representing a high risk for human health.

Hepatic enzyme induction and mutagenicity of airborne particulate matter from Santiago, Chile in the nourished and malnourished rat.

1. Respirable, airborne particles in the ambient air in downtown Santiago, Chile, have been characterized for the seasonal variation in total polycyclic aromatic hydrocarbon content, 13 of which have been identified including the mutagens (benzo(a)pyrene, dibenzo(a,h)anthracene, benzo(a)anthracene, benzo(b)fluoranthene and indeno(1,2,3, c,d)pyrene amongst others. 2. Organic extracts derived from these particles were administered to both the nourished and malnourished rat and resulted in modulation of the hepatic mixed function oxidase system including induction of NADPH-cytochrome P450 reductase, cytochrome P4501A as determined by Western blot analysis and the associated ethoxyresorufin O-deethylase and aryl hydrocarbon hydroxylase activities. 3. The cytochrome P4504A1-dependent 12-hydroxylation of lauric acid was induced in the malnourished state, but this activity was significantly inhibited by treatment of the animals with particle extracts in both nutritional states. 4. The particle extracts contained both direct and indirect-acting mutagens in the Ames test, and depending on the relative complement of both, resulted in either increased or decreased mutagenicity in the presence of S9 activation systems derived from both nourished and malnourished animals. 5. These results are discussed in the context of the interindividual risk assessment of airborne, particulate matter to man.

[Impact of outdoor pollution on indoor air quality. The case of downtown Santiago (Chile)]. / Influencia de la contaminación atmosférica en la calidad de aire de interiores. El caso del centro de Santiago (Chile).

The influence of outdoor pollution on indoor air quality was studied in downtown Santiago(Bandera street). Carbon monoxide (CO), nicotine, particulate matter, respirable fraction (PM5) and total and carcinogenic polyaromatic hydrocarbons (PAHs) were simultaneously monitored indoors and outdoors in restaurants, offices and other places. The levels of CO changed simultaneously outdoors and indoors (r = 0.89) specially during traffic rush hours, demonstrating the importance of outdoor infiltration into the indoor air quality and masking the contribution of other CO indoor sources. The maximum CO concentrations were over 800% and over 1000% higher indoors and outdoors respectively than the 9 ppm CO National Ambient Air Quality. The PM 5 concentrations were very high and showed no significant differences (p > 0.05) from indoors to outdoors, or between indoor levels in restaurants, offices and other places. Total and carcinogenic PAHs levels were also very high outdoors and indoors, outdoor levels being generally higher than those indoors and no significant; differences (p > 0.05) were found for the indoor levels between restaurants, offices and other places. Nicotine levels showed significant differences (p < 0.05) between indoor and outdoor levels. In addition, great differences (p < 0.05) in indoor levels, were found between offices and restaurants, and offices and other places. Among indoor sources cigarette smoke seems to be a minor source since nicotine concentrations, being 2.3 times higher in restaurants and other places than in offices, do not contribute to enhance significantly PM5 and total and carcinogenic HAPs in the first ones. These results suggest that in downtown Santiago, infiltration might be the main source of indoor pollution. This is supported by two evidences: a) coronene, a tracer of vehicle emissions was found in high concentration indoors and b) in restaurants (in which PAHs emissions might be higher indoor) a correlation coefficient of 0.987 for the indoor and outdoor concentrations of carcinogenic PAHs was found. Furthermore a survey asking for different symptoms and effects probably related to air pollution was made to people working in Bandera and in a rural area located 40 Km from Santiago. The results showed that excluding smoking as a confoundend factor, people working in Bandera showed a significantly greater (p < 0.05) risk of ill effects on their health than people working in the rural area.

Mutagenicity of organic extracts from Santiago (Chile) airborne particulate matter.

The Ames test has been used to detect the mutagenic activity of organic extracts from Santiago (Chile) airborne particles collected in 1990 and 1991 in one of the monitoring net system stations (MACAM). The samples were assayed with the strains TA98, TA98-NR, and TA98/1,8-DNP6 of Salmonella typhimurium, in the presence and in the absence of liver S9 fraction obtained from rats treated with Aroclor 1254. With the strain TA98 all the samples showed a very high mutagenic response either in the presence or in the absence of S9 fraction, suggesting that Santiago airborne particles contain both indirect-acting (polycyclic aromatic hydrocarbons) and direct-acting mutagenic agents. The mutagenicity of Santiago airborne particles was much higher than that reported in studies performed in other countries. Results obtained with the strains TA98-NR and TA98/1,8-DNP6 suggest that the extracts also contain mononitro and dinitroarenes. These nitroarenes have been described as very potent mutagenic agents, that can be generated by photochemical reactions under certain atmospheric conditions, or in the combustion of fuel, especially of diesel motors. The presence of polycyclic aromatic hydrocarbons and nitroarenes in Santiago airborne particles, as well as the high levels of mutagenicity detected, suggest that the inhabitants permanent exposure to these kinds of compounds represents a high risk for human health.