Cytochrome P450 1A1 induction in rat lymphoid tissues following in vivo and in vitro exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin requires protein kinase C.
The induction of cytochrome P450 1A1 (CYP1A1) is one of the most sensitive responses associated with exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related compounds. The mechanisms that underlie this response are not completely understood, particularly in lymphoid tissues that may be used in biomarker studies in humans. CYP1A1 mRNA expression and enzyme activity (ethoxyresorufin-o-deethylase, EROD) were investigated in rat thymus and spleen and isolated thymocytes and splenocytes in culture. Thymus- or spleen-derived microsomes from rats treated in vivo with TCDD showed induced EROD activity after as little as 12 h following a single exposure to TCDD (5 microg/kg body weight). Resting rat thymocytes in culture had detectable levels of EROD activity and CYP1A1 mRNA expression which increased following in vitro exposure to > or = 0.1 nM TCDD for 24 or 48 h. Interestingly, concomitant in vitro exposure of rat thymocytes to TCDD and the mitogen concanavalin A (Con A) inhibited the induction of EROD activity, which is in contrast to the response of cultured human peripheral blood lymphocytes (Landi et al., 1994; Pharmacogenetics 4, 242 246). Resting rat splenocytes in culture had no detectable EROD activity and CYP1A1 activity could not be induced by in vitro TCDD exposure, in the presence or absence of Con A. These results suggest that the relative maturation state of the cells is important in regulating the expression of CYP1A1, since splenocytes represent a more mature population of B and T lymphocytes. TCDD-induced CYP1A1 expression in cultured rat thymocytes was inhibited by the addition of calphostin C, a specific protein kinase C (PKC) inhibitor, suggesting a role for PKC as a second messenger in the CYP1A1 induction pathway. In vivo co-exposure with phorbol-myristate-acetate (PMA) and TCDD also inhibited CYP1A1 induction. Again, suggesting a role for PKC in CYP1A1 induction. Together, these results indicate that relative lymphocyte maturation state and the PKC pathway are important factors in regulating the expression of CYP1A1.
Stephen
F D
FD
Department of Pharmacology and Toxicology, State University of New York at Buffalo, 14214, USA.
Drahushuk
A T
AT
Olson
J R
JR
D000818:Animals; D019363:Cytochrome P-450 CYP1A1 / Q000096:biosynthesis* / Q000235:genetics; D004790:Enzyme Induction / Q000187:drug effects; D008221:Lymphoid Tissue / Q000187:drug effects* / Q000201:enzymology; D008297:Male; D000072317:Polychlorinated Dibenzodioxins / Q000633:toxicity*; D011493:Protein Kinase C / Q000502:physiology*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000072317:Polychlorinated Dibenzodioxins; D012333:RNA, Messenger; D019363:Cytochrome P-450 CYP1A1; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate