Gene expression of normal human epidermal keratinocytes modulated by trivalent arsenicals.

Chronic exposure to inorganic arsenic (iAs) is associated with the development of benign and malignant human skin lesions including nonmelanoma skin cancers. The precise arsenical form(s) responsible for this carcinogenic effect are unknown, although trivalent inorganic arsenic (iAs(III)) and two of its toxic metabolites, monomethylarsonous acid (MMA(III)) and methylarsinous acid (DMA(III)), are attractive candidates. In an effort to better understand and compare their toxic effects in the skin, we compared the global gene expression profiles of normal human epidermal keratinocytes (NHEKs) exposed to varying noncytotoxic/slightly cytotoxic concentrations of iAs(III), MMA(III), and DMA(III) for 24 h. Exposure to each arsenical treatment group exhibited a dose effect in the number of altered genes and the magnitude of expression change in NHEKs. The most significant gene expression changes associated with iAs(III) and MMA(III) exposure were consistent with several key events believed to be important to As-driven skin carcinogenesis, namely induction of oxidative stress, increased transcript levels of keratinocyte growth factors, and modulation of MAPK and NF-κB pathways. At both comparable arsenical concentrations and comparable NHEK toxicity, greater potential carcinogenic effects were observed in MMA(III)-exposed NHEKs than those exposed to iAs(III), including involvement of more proinflammatory signals and increased transcript levels of more growth factor genes. In contrast, none of these above-mentioned transcriptional trends were among the most significantly altered functions in the DMA(III) treatment group. This study suggests the relative capacity of each of the tested arsenicals to drive suspected key events in As-mediated skin carcinogenesis is MMA(III) > iAs(III) with little contribution from DMA(III).

Global gene expression profiling of hyperkeratotic skin lesions from inner Mongolians chronically exposed to arsenic.

The skin is an organ that is highly sensitive to chronic arsenic (As) exposure. Skin lesions such as hyperkeratoses (HKs) are common early manifestations of arsenicosis in humans. HKs can be precursor lesions of nonmelanoma skin cancers (NMSCs), but the driving forces behind their formation and how they may ultimately progress to NMSCs are unknown. The goal of this study was to examine the global gene expression profiles of As-related HKs in an effort to better understand gene expression changes that are potentially associated with early stages of As carcinogenesis. HK biopsies were removed from individuals living in an arsenicosis-endemic region in Inner Mongolia who had been exposed to high As levels in their drinking water for >20 years. Gene expression profiling was performed on RNA isolated from 7 individuals in this group and from 4 lesion-free skin samples from healthy individuals. Consistent with the pathological characteristics of the HK lesions, major functional categories and known canonical pathways represented by altered transcripts include those involved in development, differentiation, apoptosis, proliferation, and stress response. The results of this study may help define a signature profile of gene expression changes associated with long-term As exposure in the skin.

Toxicoproteomics and its application to human health risk assessment.

Toxicoproteomics is the use of proteomic technologies to better understand environmental and genetic factors, toxic mechanisms, and modes of action in response to acute exposure to toxicants and in the long-term development of diseases caused or influenced by these exposures. Use of toxicoproteomic technologies to identify key biochemical pathways, mechanisms, and biomarkers of exposure and toxicity will decrease the uncertainties that are associated with human health risk assessments. This review provides an overview of toxicoproteomics from human health risk assessment perspectives. Key toxicoproteomic technologies such as 2-D gel-based proteomic methods and toxicoproteomic approaches are described, and examples of applications of these technologies and methodologies in the risk assessment context are presented. The discussion includes a focus on challenges and future directions.

Kidney toxicogenomics of chronic potassium bromate exposure in f344 male rats.

BACKGROUND: Potassium bromate (KBrO3), used in both the food and cosmetics industry, and a drinking water disinfection by-product, is a nephrotoxic compound and rodent carcinogen. To gain insight into the carcinogenic mechanism of action and provide possible biomarkers of KBrO3 exposure, the gene expression in kidneys from chronically exposed male F344 rats was investigated. METHODS: Male F344 rats were exposed to KBrO3 in drinking water for 52 and 100 wk. Kidneys were removed, frozen, and stored at -80°C, then used for Affymetrix microarray analysis. Gene expression patterns were examined using a non-carcinogenic (20 ppm) and carcinogenic dose (400 ppm) at 52 wk, and compared to 100 wk high dose (400 ppm) and adenoma gene expression. RESULTS: Statistical analysis revealed 144, 224, 43, and 994 genes out of 15866 from the 52 wk low, 52 wk high, 100 wk high, and adenomas respectively, were differentially expressed when compared to control kidneys. Gene ontology classification of the 52 wk high dose showed alterations of gene transcripts involved in oxidative stress, lipid metabolism, kidney function/ion transport, and cellular function. In a comparison of kidney development gene expression, alterations were seen in the adenomas but not in the 52 wk bromate-treated kidneys. However, the normal kidney from the high dose group resembled the adenoma expression pattern with early kidney development genes being up-regulated and adult phase genes being down-regulated. Moreover, eight genes were identified which could serve as biomarkers of carcinogenic exposure to bromate. The most promising of these was Pendrin, or Slc26a4, a solute carrier of chloride and iodide active in the kidney, thyroid, and inner ear. All these tissues are targets of KBrO3 toxicity. Expression array results were verified with quantitative real-time rtPCR. CONCLUSIONS: These data demonstrate that the 400 ppm carcinogenic dose of KBrO3 showed marked gene expression differences from the 20 ppm non-carcinogenic dose. Comparison of kidney development gene expression showed that the adenoma patterns were more characteristic of embryonic than adult kidneys, and that the normal kidney from the high dose group resembled the adenoma-like gene expression pattern. Taken together, the analysis from this study identifies potential biomarkers of exposure and illuminates a possible carcinogenic mode of action for KBrO3.

The effect of arsenicals on ultraviolet-radiation-induced growth arrest and related signaling events in human keratinocytes.

The molecular mechanisms mediating arsenic-induced carcinogenesis are not well understood. The role of confounding factors such as ultraviolet radiation (UV), add another level of complexity to the study of arsenic carcinogenesis and the cancer-risk assessment on humans. We hypothesized that arsenicals are capable of overriding the growth arrest caused by UV treatment and may lead to selective proliferation. To test this hypothesis, a primary normal human epidermal keratinocyte (NHEK) cell culture model was used. One group was pre-exposed to UVB (100 mJ/cm(2)) that arrested a majority ( approximately 95%) of cells in G0/G1 (+UV) and a second group was not exposed to UV (-UV). Treatment of cells with various arsenicals [0-12 microM of inorganic arsenite (iAs), 0-2 microM of methyl oxoarsine (MMAs III) and 0-3 microM of iododimethyl arsine (DMAs III)] indicated a concentration-dependent increase in proliferation at 24 h in the order of DMAs III > MMAs III > iAs. Flow-cytometric analyses revealed differential effects on cell cycle distribution. Analysis of a battery of cell cycle proteins (cyclin D1, cdk5, PCNA, cdc25A and cdc25C) indicated exposure-specific differential expression profiles. Increased activation of JNK phosphorylation (5-10-fold) in the +UV group and the synergistic increase with methyl arsenicals suggested that JNK might be involved in cell survival and proliferative signaling. Induction of EGF levels and increased phosphorylation of the EGF receptor by arsenicals (+UV) suggested that the EGF signaling pathway might mediate arsenical-induced cell proliferation of NHEK cells. Differential activation of ERK1/2 by arsenicals (+/-UV) suggested that EGF-mediated cell proliferation by arsenicals in UV-treated NHEK cells may not involve ERK activation. Taken together, the data suggest that both UV exposure and methylation status of the arsenicals dictate the participation of key cell cycle proteins and related signaling events in arsenical-induced cell proliferation.

Non-thermal exposure to radiofrequency energy from digital wireless phones does not affect ornithine decarboxylase activity in L929 cells.

L929 murine fibroblast cells were exposed to radiofrequency (RF) radiation from a time division multiple access wireless phone operating at 835 MHz frequency to determine the effect of RF-radiation energy emitted by wireless phones on ornithine decarboxylase (ODC) activity in cultured cells. Exposure was for 8 h to an average specific absorption rate (SAR) from <1 W/kg up to 15 W/kg. After exposure, cells were harvested and ODC activity was measured. No statistically significant difference in ODC activity was found between RF-radiation-exposed and sham-exposed cells at non-thermal specific absorption rates. At SARs which resulted in measurable heating of the medium, a dose-dependent decrease in enzymatic activity was observed and was shown to be consistent with a comparable decrease caused by non-RF-radiation heating. Thus we observed only the well-known enzyme inhibition due to heating, rather than the previously reported enhancement attributed to RF-radiation exposure.