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Comparison of expression profiles of drug‐metabolizing enzymes between human liver and the hepatic cell lines HepG2, Hep3B, and HuH7: a parallel reaction monitoring (PRM)‐based targeted proteomics study
Author(s) -
Shi Jian,
Wang Xinwen,
Zhu HaoJie
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.668.9
Subject(s) - drug metabolism , enzyme , isozyme , proteomics , cell culture , microsome , quantitative proteomics , in vitro , chemistry , biochemistry , biology , microbiology and biotechnology , gene , genetics
Background Several human hepatic cell lines have been widely used as an in vitro model for drug metabolism related studies because of their availability and immortalization. However, the validity of this model is a subject of debate as the expression patterns of drug‐metabolizing enzymes (DMEs) in these cells could differ significantly from that of human livers. In the present study, we systematically quantified the basal protein expression of 61 clinically important DMEs in the human liver‐derived cell lines HepG2, Hep3B, and HuH7, and compared the expression profiles to that in human livers. Methods With high resolution and high mass accuracy, parallel reaction monitoring (PRM) is a new powerful quantitative proteomics tool, providing wider dynamic range and higher selectivity than selected reaction monitoring (SRM). A PRM‐based targeted proteomics assay was developed for relative quantification of DMEs in the microsomes prepared from human livers and the hepatic cell lines HepG2, Hep3B, and HuH7. The 45 targeted enzymes include 14 cytochromes P450 enzymes (CYPs), 9 UDP‐glucuronosyltransferases (UGTs), and 38 other enzymes relevant to clinical drug metabolism. Results With the exception of aldehyde dehydrogenase 1 family member A1 (ALDH1A1), glutathione s‐transferase omega‐1 (GSTO1), and catechol‐O‐methyltransferase (COMT), most of the DMEs are expressed at a substantially lower level relative to that in human livers. In addition, the expression levels of different enzymes vary markedly among the three hepatic cell lines. Conclusions Protein expression profiles of DMEs differ significantly between human livers and the hepatic cell lines HepG2, Hep3B, and HuH7. Appropriate caution must be exercised when considering these cell lines for drug metabolism studies, and interpreting the data from the studies based on these cell lines.