Identification of cytochromes P450 1A2, 2A1, 2C7, 2E1 in rat glioma C6 cell line by RT-PCR and specific restriction enzyme digestion.
Reverse transcription (RT)/polymerase chain reaction (PCR) was used to detect the expression of six isoforms of cytochrome P450 which belong to five P450 subfamilies in rat glioma C6 cell line. P450 1A2, 2A1, 2C7, 2E1 were identified by RT-PCR in all samples, including untreated cells as well as cells treated with phenobarbital (PB) or benzo(a)anthracene (BA). P450 3A and 2C11 were not detected in our glioma samples although they were detected in liver tissues in our previous study. To confirm proper PCR products, various restriction enzymes were used to digest the PCR fragments and the expected digestion patterns were obtained. These results demonstrate for the first time that glioma C6 cells, representing a single cell type of rat central nervous system (CNS), contain a P450-dependent metabolism system which seems important for understanding drug metabolism, neurotransmission as well as tumor etiology and chemotherapy in brain.
Geng
J
J
Department of Biochemistry and Molecular Biology, University of Texas Medical School, Houston 77225.
Strobel
H W
HW
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D001483:Base Sequence; D001551:Benz(a)Anthracenes / Q000494:pharmacology; D001921:Brain / Q000187:drug effects / Q000201:enzymology*; D002460:Cell Line; D019388:Cytochrome P-450 CYP1A2; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis / Q000096:biosynthesis*; D017931:DNA Primers; D004790:Enzyme Induction; D005910:Glioma / Q000201:enzymology* / Q000235:genetics; D008297:Male; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000032:analysis / Q000096:biosynthesis*; D010634:Phenobarbital / Q000494:pharmacology; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D012194:RNA-Directed DNA Polymerase; D051381:Rats; D017207:Rats, Sprague-Dawley; D015183:Restriction Mapping; D013250:Steroid Hydroxylases / Q000032:analysis / Q000096:biosynthesis*; D014407:Tumor Cells, Cultured
D001551:Benz(a)Anthracenes; D017931:DNA Primers; D012333:RNA, Messenger; D003577:Cytochrome P-450 Enzyme System; C030935:benz(a)anthracene; D010088:Oxidoreductases; D013250:Steroid Hydroxylases; D001189:Aryl Hydrocarbon Hydroxylases; D019388:Cytochrome P-450 CYP1A2; C029009:steroid hormone 7-alpha-hydroxylase; D012194:RNA-Directed DNA Polymerase; D010634:Phenobarbital