Effects of tobacco compounds on gene expression in fetal lung fibroblasts.
The goal of this study was to determine the effects of tobacco compounds on gene expression in a human fetal lung cell line (WI38). In the present study, we investigated the effects of tobacco compounds (nicotine, benzo(a)pyrene (B(a)P), and 2-Naphthylamine) on gene expression profiles in human fetal fibroblasts using cDNA microarray and real-time PCR. WI38 cells were cultured in Eagle's minimum essential medium (MEM) supplemented with 10% fetal bovine serum, 2% 200 mM L-glutamine, and a 2% penicillin and streptomycin solution. Tissue culture flasks (T-25 cm(2)) containing confluent lung fibroblasts were incubated at 37 degrees C for 24 h with 5 mL of medium supplemented with 10 microM of a tobacco compound (nicotine, B(a)P, or 2-Naphthylamine). The gene expression profiles for the W138 cells varied depending on the tobacco compound. The cDNA microarray analysis revealed that apoptosis-related genes such as DNASE2, MADD, MST1, NME3, RARG, TNFRSF1A, BAD, and DFFB genes were down-regulated in tobacco compound-treated WI38 cells. We also observed significant increases in Arnt gene expression by real-time PCR in tobacco compound-treated WI38 cells. Tobacco compounds can affect apoptosis, immunity, and growth in WI38 cells. A microarray-based genomic survey is a high-throughput approach for the evaluation of gene expression in cell lines treated with tobacco compounds.
Sohn
Sung-Hwa
SH
Department of Biochemistry & Molecular Biology, College of Medicine, Korea University, Seoul 136-705, Korea.
Kim
Ki-Nam
KN
Kim
In Kyoung
IK
Lee
Eun-Il
EI
Ryu
Jae-Jun
JJ
Kim
Meyoung-Kon
MK
D015081:2-Naphthylamine / Q000032:analysis / Q000494:pharmacology; D001564:Benzo(a)pyrene / Q000032:analysis / Q000494:pharmacology; D002460:Cell Line; D002470:Cell Survival; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008168:Lung / Q000166:cytology* / Q000196:embryology; D009538:Nicotine / Q000032:analysis / Q000494:pharmacology; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014026:Nicotiana / Q000737:chemistry*
D001564:Benzo(a)pyrene; D009538:Nicotine; D015081:2-Naphthylamine