Stable expression of cytochrome P450IIIA7 cDNA in human breast cancer cell line MCF-7 and its application to cytotoxicity testing.
A mammalian cell expression plasmid containing cytochrome P450IIIA7 complementary DNA was constructed. Breast cancer cells (MCF-7) were transfected with the plasmid and neomycin-resistant selection marker plasmid. We established three cell lines, termed M13, M21, and M27, which expressed the cytochrome P450IIIA7 as examined by RNA blot and immunoblot analyses. These cell lines showed 8- to 10-fold higher sensitivity against aflatoxin B1 compared to parental MCF-7 cells, suggesting that cytochromes P450IIIA7 expressed in the cells were responsible for the production of the cytotoxic metabolite of aflatoxin B1.
Kitamura
R
R
Division of Analytical Biochemistry, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
Sato
K
K
Sawada
M
M
Itoh
S
S
Kitada
M
M
Komori
M
M
Kamataki
T
T
D000328:Adult; D016604:Aflatoxin B1 / Q000633:toxicity*; D001483:Base Sequence; D001943:Breast Neoplasms / Q000188:drug therapy / Q000201:enzymology / Q000235:genetics*; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics; D004273:DNA, Neoplasm / Q000737:chemistry*; D004354:Drug Screening Assays, Antitumor; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D015151:Immunoblotting; D008099:Liver / Q000196:embryology / Q000201:enzymology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D004273:DNA, Neoplasm; D003577:Cytochrome P-450 Enzyme System; D016604:Aflatoxin B1