Transcript profiling of cytochrome P450 genes in HL-60 human leukemic cells: upregulation of CYP1B1 by all-trans-retinoic acid.
All-trans-retinoic acid (ATRA) is used in the treatment of promyelocytic acute leukemia. The biotransformation of this drug is catalyzed by various cytochrome P450 (CYP) enzymes, but relatively little is known about the effect of ATRA on CYP enzyme expression in leukemic cells. In the present study, we conducted transcript profiling of CYP and related genes in cultured HL-60 human promyelocytic leukemic cells and determined the effect of ATRA on the expression of these genes. Reverse transcription-polymerase chain reaction (RT-PCR) analysis with a block-cycler indicated the presence of CYP1B1 but not CYP1A1, CYP2B6, CYP2C8, CYP2C9, CYP3A4, CYP3A5, or CYP26A1 transcript in cultured HL-60 cells. ATRA treatment (0.1-40 microM for 3 days) increased CYP1B1 mRNA levels by up to 3 fold, as determined by a quantitative real-time PCR method. The same ATRA treatment also resulted in the detection of CYP26A1 but not CYP1A1, CYP2B6, CYP2C8, CY2C9, CYP3A4, or CYP3A5 mRNA. Additional experiments showed that phenobarbital increased CYP2B6 mRNA expression and that pregnane X receptor (PXR) but not constitutive androstane receptor (CAR) was detected in HL-60 cells. Overall, our novel findings indicate the upregulation of CYP1B1 by ATRA in HL-60 human promyelocytic leukemic cells shown for the first time to express PXR but not CAR mRNA.
Kawai
Masahiko
M
Faculty of Pharmaceutical Sciences, The University of British Columbia, Vancouver, BC, Canada.
Chen
Jie
J
Cheung
Catherine Y S
CY
Chang
Thomas K H
TK
D001189:Aryl Hydrocarbon Hydroxylases / Q000096:biosynthesis*; D000090702:Constitutive Androstane Receptor; D065633:Cytochrome P-450 CYP1B1; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000235:genetics*; D018076:DNA, Complementary / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D000077297:Pregnane X Receptor; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D011987:Receptors, Steroid / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic; D014212:Tretinoin / Q000378:metabolism*; D015854:Up-Regulation
D000090702:Constitutive Androstane Receptor; D018076:DNA, Complementary; D000077297:Pregnane X Receptor; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; D011987:Receptors, Steroid; D014157:Transcription Factors; D014212:Tretinoin; D012313:RNA; D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; C585496:CYP1B1 protein, human; D065633:Cytochrome P-450 CYP1B1