Oxidative stress induces GSTP1 and CYP3A4 expression in the human erythroleukemia cell line, K562.
To explore the effects of oxidative stress on metabolizing enzymes in blood cells, we examined the effect of hydrogen peroxide (H(2)O(2)) on glutathione S-transferase (GST) and cytochrome P450 (CYP) expression in a human erythroleukemic cell line, K562. After adding H(2)O(2) (up to 100 microM) to the culture medium of K562 cells, the expression levels of GST and CYP enzymes were monitored by RT-PCR. The expression of GSTP1 and CYP3A4 was induced by oxidative stress. Quantitative PCR and immunoblot analysis revealed a 3- to 4-fold increase in GSTP1 and CYP3A4 mRNA, and a 2- to 3-fold increase in GSTP1 and CYP3A4 protein levels, 3 d after the addition of 100 microM H(2)O(2). Induction was H(2)O(2) dose-dependent and also depended on the length of culture. Our results suggest that oxidative stress may affect GST and/or CYP expression in human blood cells, which may alter the metabolism of drugs and xenobiotics and thus, the toxicity of these compounds to the blood cells.
Nagai
Fusako
F
Kyoritsu University of Pharmacy, Shibakoen, Minato-ku, Tokyo 105-8512, Japan.
Kato
Eriko
E
Tamura
Hiro-omi
HO
D002455:Cell Division / Q000187:drug effects; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000235:genetics; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D020014:K562 Cells; D016877:Oxidants / Q000633:toxicity; D018384:Oxidative Stress; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors
D016877:Oxidants; D003577:Cytochrome P-450 Enzyme System; D006861:Hydrogen Peroxide; C104464:CYP3A protein, human; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D005982:Glutathione Transferase