Evaluation of human CYP1A2 and CYP3A4 mRNA expression in hepatocytes from chimeric mice with humanized liver.
We investigated and compared the expression of human CYPs mRNA in primary cultures of cryopreserved human hepatocytes and in chimeric mice constructed by transplanting hepatocytes from the same human donors. Analysis was performed by real-time reverse-transcription polymerase chain reaction. Initial expression levels for the 12 human CYPs mRNA in chimeric mouse hepatocytes were higher than those in human hepatocytes, but a low correlation coefficient was observed (r=0.690). After 24 h of culture, the correlation remained low (r=0.699). The medium was replaced with fresh medium without human epidermal growth factor, and after 48 h of culture, expression of the 12 human CYPs mRNA were very similar in human hepatocytes and chimeric mouse hepatocytes, and a higher correlation coefficient was observed (r=0.809). After 72 h of culture, the correlation remained high (r=0.873). The ratio of human CYP1A2 mRNA to beta-actin mRNA in chimeric mouse hepatocytes decreased quickly during the first 24 h of culture, and then remained constant. Expression profiles of human CYP1A2 mRNA in chimeric mouse hepatocytes were similar to those in human hepatocytes after exposure of beta-naphthoflavone. CYP3A4 mRNA expression was increased significantly by rifampicin (Rif) exposure in human hepatocytes, whereas Rif-induced increases in CYP3A4 mRNA expression in chimeric mouse hepatocytes was seen for two of the three donors. In conclusion, we demonstrated that expression and induction of human CYPs in human hepatocytes can be reproduced in chimeric mouse hepatocytes.
Yoshitsugu
Hiroki
H
Division of Pharmacology, Drug Safety and Metabolism, Otsuka Pharmaceutical Factory, Inc., Naruto, Tokushima.
Nishimura
Masuhiro
M
Tateno
Chise
C
Kataoka
Miho
M
Takahashi
Eiji
E
Soeno
Yoshinori
Y
Yoshizato
Katsutoshi
K
Yokoi
Tsuyoshi
T
Naito
Shinsaku
S
D000818:Animals; D017690:Cell Transplantation; D002478:Cells, Cultured; D019388:Cytochrome P-450 CYP1A2 / Q000235:genetics*; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology* / Q000637:transplantation; D006801:Humans; D008297:Male; D051379:Mice; D016513:Mice, SCID; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012293:Rifampin / Q000494:pharmacology; D013997:Time Factors; D018183:Transplantation Chimera / Q000235:genetics*; D014183:Transplantation, Heterologous; D019324:beta-Naphthoflavone / Q000494:pharmacology
D012333:RNA, Messenger; D019324:beta-Naphthoflavone; D003577:Cytochrome P-450 Enzyme System; C504573:CYP1A2 protein, human; D019388:Cytochrome P-450 CYP1A2; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D012293:Rifampin