Low-level arsenite induced gene expression in HEK293 cells.

TitleLow-level arsenite induced gene expression in HEK293 cells.
Publication TypeJournal Article
Year of Publication2003
AuthorsZheng XHui, Watts GS, Vaught S, A Gandolfi J
Date Published2003 May 01
KeywordsArsenites, Cell Line, DNA, Complementary, Dose-Response Relationship, Drug, Down-Regulation, Gene Expression Profiling, Gene Expression Regulation, Heme Oxygenase (Decyclizing), Humans, Multigene Family, Oligonucleotide Array Sequence Analysis, Oxidative Stress, Reverse Transcriptase Polymerase Chain Reaction, Sodium Compounds, Up-Regulation

Chronic, low-level exposure to arsenic frequently results in skin, lung, bladder, and kidney cancer. Since arsenic is primarily excreted via the kidney, this study focused on this target tissue. Gene array was used as a sensitive low-level monitor of the impact of arsenic on this target tissue. Arsenite [As(III)] was chosen as the chemical species of arsenic since As(III) species are touted as the cellular toxic form of arsenic. Human embryonic kidney cell line HEK293 cells were incubated with 1, 10, and 25 microM arsenite [As(III)] for 6 or 24 h. Total RNA from treated and control cells was isolated, reverse transcribed, and labeled with Cy3 or Cy5, and hybridized to a human cDNA microarray. Hybridizations were performed four times using independent total RNA preparations to ensure reproducibility. Raw data from 10 and 25 microM treated cells exposed for 6 h was normalized within, and between, hybridizations followed by identification of genes affected by arsenite exposure based on practical significance (2-fold change up or down) and reproducibility (affected in four of six measurements). In these studies, 20 genes (HMOX1, MT1E, or FOSL1, etc.) were up-regulated, and 19 genes (MYC, JAK1, or CENPE, etc.) were down-regulated. Genes identified at 10 and 25 microM arsenic exposure were then examined after 1 microM treatment for 6 or 24 h. Expression of affected genes showed a dose-dependent (1-25 microM) trend that was apparently not time-dependent (6 vs. 24 h). The affected genes indicate that even this realistic, low-level arsenite exposure was recognized by the HEK293 cells (e.g. metallothionein genes) and produced an oxidative stress (e.g. heme oxygenase gene). These affected genes were characterized as stress response genes, proto-oncogene, signaling molecules, transcription factors, chemokine receptors, proteolytic enzymes, ESTs, and unknown genes. These findings imply that arsenite induces complex cellular injury and the cellular adaptation to As(III) is associated with alterations in the expression of many genes.

Alternate JournalToxicology
PubMed ID12679051
Grant ListP30 CA023074 / CA / NCI NIH HHS / United States
04940 / / PHS HHS / United States
P30 ES006694 / ES / NIEHS NIH HHS / United States
06694 / / PHS HHS / United States
CA 023074 / CA / NCI NIH HHS / United States