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Identification of Genes Responding to Low-Dose Arsenite Using HiCEP

ABSTRACT Chronic ingestion of arsenic in polluted food and water can cause various human disorders including skin and lung cancers. Sensitive biomarkers from human tissue/cells could help to prevent chronic intoxication with low-dose arsenite. Using High-Coverage Expression Profiling (HiCEP), an Amplified Fragment Length Polymorphism (AFLP)-based gene expression profiling technique, we analyzed the expression of approximately 11,000 genes in human lung fibroblasts (HFLIII) and compared the profiles between cells, treated and untreated with 1 μM sodium arsenite (NaAsO2). Hundreds of genes appeared upregulated and downregulated more than two-fold, 2 h after the treatment. Marked induction was found (>4.4-fold) in a few genes including HMOX1, INHBA, and ANKRD11. Induction of the HMOX1 was detected with a dose of arsenite at as low as 0.3 μM (0.04 ppm) and reached its maximum at 4 h after the treatment. The arsenite-induced HMOX1 expression was attenuated by the promoted glutathione (GSH) synthesis by N-acetyl-L-cysteine (NAC). However, it was not affected by pretreating the cells with general radical scavengers, consistent with the fact that ionizing radiation at either high-or low-doses has never induced HMOX1 in the same assay system. Thus, induction of HMOX1 gene is highly sensitive and also selective against arsenite in the cells. The present process could provide a useful strategy for exploring biomarkers that might help in assessing the known and unknown risks of any natural and artificial toxic reagents.