Butein up-regulates the expression of the π class of glutathione S-transferase in rat primary hepatocytes through the ERK/AP-1 pathway.
Induction of phase II enzymes is an important mechanism of chemoprevention. Here we compared the effects of chalcones on the expression of the π class of glutathione S-transferase (GSTP) in rat primary hepatocytes. Hepatocytes were treated with 10 or 25 μM of phloretin or butein for 24 h. Both butein and phloretin dose-dependently increased GSTP protein expression, and the induction potency of butein was stronger than that of phloretin. The increase in GSTP mRNA in cells treated with 25 μM of phloretin and butein was 107% and 211%, respectively (P < 0.05). Butein increased GST enzyme activity by 27% compared with that in the control cells (P < 0.05). In contrast, phloretin had no significant effect on GST enzyme activity. The pTA-luciferase reporter construct with the rat -2.7 kb GSTP promoter region was transiently transfected into Clone 9 liver cells, and the luciferase activity in butein-treated cells was 1.1-fold higher than that in control cells (P < 0.05). GSTP enhancer 1 (GPE1) deletion abolished the induction of reporter activity by butein. The phosphorylation of extracellular signal-regulated kinase (ERK), but not of c-Jun NH2-terminal kinase (JNK) and p38, was stimulated in the presence of butein. Pretreatment with PD98059, an ERK inhibitor, alleviated the increase in activator protein-1 (AP-1)-DNA binding activity and also the activation of GSTP protein expression by butein. Moreover, c-Jun is likely to bind to the GPE1. Silencing of ERK2 by siRNA gene knockdown reduced the butein-induced expression of GSTP. In conclusion, the increased GSTP expression by butein is likely related to the ERK-AP-1 pathway.
Chang
Hebron C
HC
Department of Biotechnology, Asia University, NO. 500, Lioufeng Road, Wufeng, Taichung 413, Taiwan.
Chen
Haw-Wen
HW
Tung
Hui-Shan
HS
Liu
Kai-Li
KL
Tsai
Chia-Wen
CW
Lii
Chong-Kuei
CK
D000818:Animals; D047188:Chalcones / Q000494:pharmacology*; D024202:Electrophoretic Mobility Shift Assay; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism*; D022781:Hepatocytes / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D008297:Male; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D015854:Up-Regulation
D047188:Chalcones; D012333:RNA, Messenger; D018808:Transcription Factor AP-1; C040918:butein; D005982:Glutathione Transferase; D048049:Extracellular Signal-Regulated MAP Kinases