|
KMID : 0617319980070010144
|
|
Journal of Pharmacetical Sceiences Ewha Womans University
1998 Volume.7 No. 1 p.144 ~ p.149
|
|
|
Stimulation of Trout CYP1A Gene Expression in Mouse HEPA-1 Cells by 3-Methylcholanthrene
|
|
Lee Soo-Young
Sheen Yhun-Yhong
|
|
|
Abstract
|
|
|
|
Trout CYP1A-CAT expression construct was generated by cloning-3.5kb 5´ flanking DNA of trout liver CYP1A gene in front of CAT gene at pCAT-basic vector. Hepa 1 cells, which an known to contain a functional arylhydrocarbon receptor´ were transfected with trout CYP1A CAT using lipofectin. 3-Methylcholanthrene (1 nM) was added into hepa 1 cells in culture in order to examine if 5´ flanking DNA of trout CYP1A gene could interact with mouse transactivating factors to bring about transcription of the chloramphenicol acetyltransferase(CAT) reporter gene. The level of CAT protein was measured by CAT ELISA and the level of CAT mRNA was determined by RTPCR. The treatment of 1 nM 3-methylcholanthrene resulted two fold increases in CAT protein as well as CAT mRNA compared to untreated control hepa cells. These data indicate that arylhydrocarbon receptors of mouse hepa 1 cells are function to activate exogenously transfected trout CYP1A-CAT construct in terms of both transcription and translation of CAT. We also examined the effect of 3-methylcholanthrene on endogeno cyp1a1 activity in hepa 1 cell. 3-Methylcholanthrene (1 nM) treatment to hepa 1 cell transfected with trout CYP1A-CAT construct stimulated the level of cyp1a1 mRNA by two folds and the activity of ethoxyresorufin-O-deethylase by two fold compared to that of contra cell. In this study we reported that trout CYP1A-CAT reporter gene expression construct could be expressed by 3-methylcholanthrene treatment in mouse hepa 1 cells. Thus trout CYP1A-CAT could serve as a good model to study the mechanism of regulation of CYP1A1 get expression.
|
|
|
KEYWORD
|
|
|
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|
|