Genetic events associated with arsenic-induced malignant transformation: applications of cDNA microarray technology.
Arsenic is a human carcinogen. Our recent work showed that chronic (>18 wk), low-level (125-500 nM) arsenite exposure induces malignant transformation in normal rat liver cell line TRL1215. In these arsenic-transformed cells, thecellular S-adenosylmethionine pool was depleted from arsenic metabolism, resulting in global DNA hypomethylation. DNA methylation status in turn may affect the expression of a variety of genes. This study examined the aberrant gene expression associated with arsenic-induced transformation with the use of Atlas Rat cDNA Expression microarrays. Poly(A(+)) RNA was prepared from arsenic-transformed cells and passage-matched control cells, and (32)P-labeled cDNA probes were synthesized with Clontech Rat cDNA Synthesis primers and moloney murine leukemia virus reverse transcriptase. The hybrid intensity was analyzed with AtlasImage software and normalized with the sum of the four housekeeping genes. Four hybridizations from separate cell preparations were performed, and mean and SEM for the expression of each gene were calculated for statistical analysis. Among the 588 genes, approximately 80 genes ( approximately 13%) were aberrantly expressed. These included genes involved in cell-cycle regulation, signal transduction, stress response, apoptosis, cytokine production and growth-factor and hormone-receptor production and various oncogenes. These initial gene expression analyses for the first time showed potentially important aberrant gene expression patterns associated with arsenic-induced malignant transformation and set the stage for numerous further studies. Mol. Carcinog. 30:79-87, 2001. Published 2001 Wiley-Liss, Inc.
Laboratory of Comparative Carcinogenesis, NIEHS, Research Triangle Park, North Carolina 27709, USA.
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Chen
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Liu
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BA
Merrick
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Waalkes
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D018053:Arsenites / Q000633:toxicity*; D018797:Cell Cycle Proteins / Q000235:genetics; D002471:Cell Transformation, Neoplastic / Q000187:drug effects* / Q000235:genetics; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis; D018076:DNA, Complementary / Q000032:analysis*; D004273:DNA, Neoplasm / Q000235:genetics*; D015870:Gene Expression; D020869:Gene Expression Profiling; D006133:Growth Substances / Q000096:biosynthesis; D022781:Hepatocytes / Q000187:drug effects* / Q000473:pathology; D006801:Humans; D007091:Image Processing, Computer-Assisted; D009363:Neoplasm Proteins / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D051381:Rats; D011916:Rats, Inbred F344; D015398:Signal Transduction / Q000235:genetics; D017670:Sodium Compounds / Q000633:toxicity*
D018053:Arsenites; D018797:Cell Cycle Proteins; D016207:Cytokines; D018076:DNA, Complementary; D004273:DNA, Neoplasm; D006133:Growth Substances; D009363:Neoplasm Proteins; D017670:Sodium Compounds; C017947:sodium arsenite