PCR-based subtraction analyses for upregulated gene transcription in cadmium-exposed rat lung type 2 epithelial cells

The aim of this study was to gain insight into early events in the lung epithelial cells following acute Cd exposure. We adopted the polymerase chain reaction (PCR)-based subtraction technique and found several genes that were upregulated in immortalized rat lung type 2 epithelial cells (SV40T2). The upregulation of those genes was confirmed by Northern blot analysis and categorized into three groups (highly, moderately, and weakly inducible genes). Heme oxygenase-1 (HO-1), HSP 72, hepatic steroid hydroxylase/CYPIIA2, and Cd-inducible gene 1 (cdig1, a new gene, Accession Nos. AB086233 and AB086234) were highly inducible genes, testosterone-repressed prostate message 2 mRNA was moderately inducible, and collagen-binding protein and cdig2 (another new gene, Accession No. AB086193) mRNAs were weakly inducible. The expression of cdig1 increased linearly with time up to 9h, while that of HO-1 reached the maximum value at 4h in response to 10 microM Cd.