Molecular effects of 1-naphthyl-methylcarbamate and solar radiation exposures on human melanocytes.

Carbaryl (1-naphthyl-methylcarbamate), a broad-spectrum insecticide, has recently been associated with the development of cutaneous melanoma in an epidemiological cohort study with U.S. farm workers also exposed to ultraviolet radiation, the main etiologic factor for skin carcinogenesis. We hypothesized that carbaryl exposure may increase deleterious effects of UV solar radiation on skin melanocytes. This study aimed to characterize human melanocytes after individual or combined exposure to carbaryl (100µM) and solar radiation (375mJ/cm2). In a microarray analysis, carbaryl, but not solar radiation, induced an oxidative stress response, evidenced by the upregulation of antioxidant genes, such as Hemeoxygenase-1 (HMOX1), and downregulation of Microphtalmia-associated Transcription Factor (MITF), the main regulator of melanocytic activity; results were confirmed by qRT-PCR. Carbaryl and solar radiation induced a gene response suggestive of DNA damage and cell cycle alteration. The expression of CDKN1A, BRCA1/2 and MDM2 genes was notably more intense in the combined treatment group, in a synergistic manner. Flow cytometry assays demonstrated S-phase cell cycle arrest, reduced apoptosis levels and faster induction of cyclobutane pyrimidine dimers (CPD) lesions in carbaryl treated groups. Our data suggests that carbaryl is genotoxic to human melanocytes, especially when associated with solar radiation.

Avaliação de melanócitos humanos expostos ao inseticida carbaril e à radiação solar em cultura / Evaluation of cultured human melanocytes exposed to carbaryl insecticide and solar radiation

O carbaril (metilcarbamato de naftila), um inseticida de amplo espectro, foi recentemente associado ao desenvolvimento de melanoma cutâneo em estudo epidemiológico de coorte com trabalhadores agrícolas norte-americanos, expostos também à radiação solar, o principal fator etiológico para o desenvolvimento de tumores cutâneos. Apesar de abrangente e bem planejado, aquele estudo epidemiológico não é suficiente para caracterizar a contribuição direta do inseticida e da radiação solar na melanomagênese. Diversos estudos têm explorado o efeito sinérgico de determinadas substâncias químicas à radiação UV, potencializando seus efeitos deletérios sobre a pele, e possivelmente contribuindo para o desenvolvimento de tumores. A hipótese deste trabalho é de que a exposição ao carbaril associada à radiação solar possa estimular a transformação de melanócitos. Esse estudo visou caracterizar melanócitos humanos após exposição individual ou combinada ao carbaril (100uM) e à radiação solar (375 mJ/ cm2). Em ensaio de microarray, o carbaril, mas não a radiação solar, induziu uma importante resposta a estresse oxidativo, evidenciada pelo aumento da expressão de genes antioxidantes, como o Hemeoxigenase-1 (HMOX1), e pela diminuição da expressão do gene MiTF, regulador da atividade melanocítica; os resultados foram confirmados por qRT-PCR. Além disso, tanto o carbaril quanto a radiação solar induziram respostas que sugerem dano ao DNA e alteração de ciclo celular. A expressão dos genes nestas categorias, como p21 e BRCA1/2, foi notavelmente mais intensa no grupo de tratamento combinado e de fato, ensaios por citometria de fluxo demonstraram parada de ciclo celular na fase S, redução do número de células em apoptose e indução mais rápida de lesões do tipo CPD neste grupo experimental. Nossos dados sugerem que o carbaril é genotóxico para melanócitos humanos, especialmente quando associado à radiação solar

Carbaryl (1-naphthyl-methylcarbamate), a broad spectrum insecticide, has recently been associated with the development of cutaneous melanoma in an epidemiological cohort study with U.S. farm workers also exposed to ultraviolet radiation, which is known to be the main etiologic factor for skin carcinogenesis. Although comprehensive and well designed, the epidemiological study is not sufficient to characterize the direct contribution of the insecticide and solar radiation in melanomagenesis. Several studies have explored the synergistic effect of certain chemicals with UV radiation, increasing its deleterious effects on the skin, possibly contributing to tumor development. We hypothesized that Carbaryl exposure associated with UV solar radiation may induce melanocyte transformation. This study aims to characterize human melanocytes after individual or combined exposure to Carbaryl (100uM) and solar radiation (375 mJ/ cm2). In a microarray analysis, Carbaryl, but not solar radiation, induced an important oxidative stress response, evidenced by the upregulation of antioxidant genes, such as Hemeoxygenase-1 (HMOX1), and downregulation of MiTF, the main regulator of melanocytic activity; results were confirmed by qRT-PCR. Moreover, both Carbaryl and solar UV induced a gene response that suggests DNA damage and cell cycle alteration. The expression of genes in these categories, such as p21 and BRCA1/2, was notably more intense in the combined treatment group in an additive manner and in fact, flow cytometry assays demonstrated cell cycle arrest in S phase, reduced apoptosis induction and faster induction of CPD lesions in this experimental group. Our data suggests that carbaryl is genotoxic to human melanocytes, especially when associated with solar radiation

Diuron-induced rat urinary bladder carcinogenesis: mode of action and human relevance evaluations using the International Programme on Chemical Safety framework.

Diuron, a high volume substituted urea herbicide, induced high incidences of urinary bladder carcinomas and low incidences of kidney pelvis papillomas and carcinomas in rats exposed to high doses (2500 ppm) in a 2-year bioassay. Diuron is registered for both occupational and residential uses and is used worldwide for more than 30 different crops. The proposed rat urothelial mode of action (MOA) for this herbicide consists of metabolic activation to metabolites that are excreted and concentrated in the urine, leading to cytotoxicity, urothelial cell necrosis and exfoliation, regenerative hyperplasia, and eventually tumors. We show evidence for this MOA for diuron using the International Programme on Chemical Safety (IPCS) conceptual framework for evaluating an MOA for chemical carcinogens, and the United States Environmental Protection Agency (USEPA) and IPCS framework for assessing human relevance.

Evaluation of diuron (3-[3,4-dichlorophenyl]-1,1-dimethyl urea) in a two-stage mouse skin carcinogenesis assay.

Diuron (3-[3,4-dichlorophenyl]-1,1-dimethyl urea) is an herbicide with carcinogenic activity in rats and mice, which have developed respectively urothelial and mammary gland tumors in long-term studies. Accordingly, diuron has been categorized as a "likely human carcinogen" by the U.S. Environmental Protection Agency. Although the carcinogenesis-initiating activity of diuron has been reported in an early initiation-promotion mouse skin study, its genotoxic potential has been disputed. It is necessary to clarify the mode of action through which it has caused rodent neoplasia and verify its relevance to humans. Herein, two experiments were developed to verify the initiating and promoting potentials of diuron in a twenty-three- and a twenty-one-week-long mouse skin carcinogenesis protocol. In one, dimethylsulfoxide (DMSO) was the solvent for the herbicide; in the other, acetone was the alternative solvent in order to verify whether DMSO had inhibitory influence on a potential cutaneous carcinogenic activity. The adopted schedule for the tumor-promoting agent 12-O-tetradecanoylphorbol 13-acetate (TPA) resulted in skin ulcers, which demonstrates the need for careful selection of TPA dose levels and frequency of application in this model. In both studies, diuron did not exert any influence on the skin carcinogenesis process, in contrast with results already reported in the literature.