Mutagenicity of bisphenol A and its suppression by interferon-alpha in human RSa cells.
Bisphenol A is used as a monomer in the production of polycarbonate plastic products. The widespread use of bisphenol A has raised concerns about its effects in humans. Since there is little information on the mutagenic potential of the chemical, the mutagenicity of bisphenol A was tested using human RSa cells, which has been utilized for identification of novel mutagens. In genomic DNA from cells treated with bisphenol A at concentrations ranging from 1x10(-7) to 1x10(-5)M, base substitution mutations at K-ras codon 12 were detected using PCR and differential dot-blot hybridization with mutant probes. Mutations were also detected using the method of peptide nucleic acid (PNA)-mediated PCR clamping. The latter method enabled us to detect the mutation in bisphenol A-treated cells at a dose (1x10(-8)M) equivalent to that typically found in the environment. Induction of ouabain-resistant (Oua(R)) phenotypic mutation was also found in cells treated with 1x10(-7) and 1x10(-5)M of bisphenol A. The induction of K-ras codon 12 mutations and Oua(R) mutations was suppressed by pretreating RSa cells with human interferon (HuIFN)-alpha prior to bisphenol A treatment. The cells treated with bisphenol A at the concentration of 1x10(-6)M elicited unscheduled DNA synthesis (UDS). These findings suggested that bisphenol A has mutagenicity in RSa cells as well as mutagens that have been tested in these cells, and furthermore, that a combination of the PNA-mediated PCR clamping method with the human RSa cell line may be used as an assay system for screening the mutagenic chemicals at very low doses.
Takahashi
S
S
Department of Biochemistry, School of Medicine, Chiba University, 1-8-1 Inohana, Chuou-ku, Chiba City, Chiba 260-8670, Japan.
Chi
X J
XJ
Yamaguchi
Y
Y
Suzuki
H
H
Sugaya
S
S
Kita
K
K
Hiroshima
K
K
Yamamori
H
H
Ichinose
M
M
Suzuki
N
N
D001358:Avian Sarcoma Viruses; D001559:Benzhydryl Compounds; D002461:Cell Line, Transformed / Q000187:drug effects / Q000201:enzymology; D003062:Codon / Q000187:drug effects; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004260:DNA Repair / Q000187:drug effects; D004351:Drug Resistance / Q000235:genetics; D004968:Estrogens, Non-Steroidal / Q000633:toxicity; D005347:Fibroblasts / Q000187:drug effects* / Q000201:enzymology; D005796:Genes / Q000187:drug effects; D011905:Genes, ras / Q000187:drug effects*; D006801:Humans; D016898:Interferon-alpha / Q000494:pharmacology*; D008861:Microsomes / Q000201:enzymology; D009152:Mutagenicity Tests; D009693:Nucleic Acid Hybridization; D010042:Ouabain / Q000494:pharmacology; D010636:Phenols / Q000037:antagonists & inhibitors / Q000633:toxicity*; D010969:Plastics / Q000737:chemistry; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013539:Simian virus 40; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics; D000098826:Bisphenol A Compounds
D001559:Benzhydryl Compounds; D003062:Codon; D003577:Cytochrome P-450 Enzyme System; D004968:Estrogens, Non-Steroidal; D016898:Interferon-alpha; D010042:Ouabain; D010636:Phenols; D010969:Plastics; D000254:Sodium-Potassium-Exchanging ATPase; C006780:bisphenol A; D000098826:Bisphenol A Compounds