Induction of cytochrome P4501A1 by autoclavable culture medium change in HepG2 cells.
1. Without the addition of xenobiotics, only by changing the culture medium can one induce extensively and transiently cytochrome P4501A1 (CYP1A1) protein and mRNA in human hepatoma HepG2 cells. The induction was aryl hydrocarbon receptor (AhR)-dependent, and was proven by: (1) the medium change activated the AhR, as judged by a electrophoretic mobility shift assay; and (2) the AhR inhibitor alpha-naphthoflavone inhibited the medium change-mediated induction. 2. Induction of CYP1A1 was related to medium prepared by autoclaving. By screening the ingredients in the medium, the serum had no effect on CYP1A1 induction, whereas both photo-oxidized and autoclaved tryptophan were shown to induce CYP1A1, as indicated by CYP1A1 protein or ethoxyresorufin-O-deethylase activity. The autoclaved tryptophan contained in an autoclavable medium was a more potent inducer of CYP1A1 than photo-oxidized tryptophan. 3. The results provide some practical suggestions with experiments related to CYP1A1.
Feng
Q
Q
Laboratory of Food and Biodynamics, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan.
Kumagai
T
T
Nakamura
Y
Y
Uchida
K
K
Osawa
T
T
D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media / Q000494:pharmacology*; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D008027:Light; D010100:Oxygen / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014364:Tryptophan / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014466:Ultraviolet Rays
D003470:Culture Media; D012333:RNA, Messenger; D014364:Tryptophan; D019363:Cytochrome P-450 CYP1A1; D010100:Oxygen