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.
Hirano
Seishiro
S
Environmental Health Sciences Division, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki 305-8506, Japan. seishiro@nies.go.jp
Kitajima
Hiroki
H
Hayakawa
Toru
T
Cui
Xing
X
Kanno
Sanae
S
Kobayashi
Yayoi
Y
Yamamoto
Megumi
M
D000818:Animals; D001483:Base Sequence; D002104:Cadmium / Q000633:toxicity*; D002461:Cell Line, Transformed; D017931:DNA Primers; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D020869:Gene Expression Profiling; D008168:Lung / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D051381:Rats; D013382:Subtraction Technique; D014158:Transcription, Genetic / Q000187:drug effects*; D015854:Up-Regulation / Q000187:drug effects*
D017931:DNA Primers; D002104:Cadmium