Study of the genetic determinants of UGT1A1 inducibility by phenobarbital in cultured human hepatocytes.
UGT1A1 is induced by phenobarbital. We investigated whether three common UGT1A1 variants are associated with the variability in UGT1A1 inducibility. Human hepatocytes were incubated with 2 mM phenobarbital for 2 and 6 days followed by 5 microM SN-38 (1 h), a UGT1A1 probe. SN-38 glucuronidation in the cell media was measured by high-performance liquid chromatography. Three UGT1A1 promoter variants [-53(TA)(6>7), -3156G > A and -3279T > G] were genotyped. Significant induction of UGT1A1 catalytic activity was observed in 82% and 100% of the cultures treated with phenobarbital for 2 days (median fold-induction = 1.6, range 1.3-2.8; n = 28) and 6 days (median fold-induction = 2.8, range 1.6-6.4; n = 16), respectively. After 2 days of treatment, a negative correlation was observed between the UGT1A1 basal activities and the fold-induction (Spearman r = -0.52, P < 0.005). By contrast, the UGT1A1 activities in the basal and induced states were highly correlated (Spearman r = 0.95, P < 0.0001). Similar results were observed after 6 days of treatment. The allele frequencies were not significantly different between induced (n = 22) and non-induced preparations (n = 6) (P > 0.05). The fold-induction was not associated with any variants (P > 0.05). The basal and induced activities were correlated with -53(TA)(6>7) (and with -3156G > A due to almost complete linkage with the -53 indel) (P = 0.001). No association was found with the -3279T > G single nucleotide polymorphism (P > 0.05). The indel at -53 affects the basal phenotype and appears to limit the hepatocyte capability of maximal induction after phenobarbital. However, variants at -53, -3156 and -3279 are not associated with variability in UGT1A1 inducibility.
Ramírez
Jacqueline
J
Department of Medicine, University of Chicago, Chicago, Illinois 60637, USA.
Komoroski
Bernard J
BJ
Mirkov
Snezana
S
Graber
Andrea Yoder
AY
Fackenthal
Donna Lee
DL
Schuetz
Erin G
EG
Das
Soma
S
Ratain
Mark J
MJ
Innocenti
Federico
F
Strom
Stephen C
SC
D000293:Adolescent; D000328:Adult; D000368:Aged; D002166:Camptothecin / Q000031:analogs & derivatives / Q000494:pharmacology; D002648:Child; D002675:Child, Preschool; D005260:Female; D014453:Glucuronosyltransferase / Q000235:genetics*; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007223:Infant; D000077146:Irinotecan; D008099:Liver / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D010597:Pharmacogenetics / Q000379:methods*; D010634:Phenobarbital / Q000494:pharmacology*; D020641:Polymorphism, Single Nucleotide; D000099148:UGT1A1 Enzyme
D002166:Camptothecin; D014453:Glucuronosyltransferase; D000077146:Irinotecan; D010634:Phenobarbital; D000099148:UGT1A1 Enzyme