タダ マサコ   Tada Masako
  多田 政子
   所属   東邦大学  理学部 生物学科
   職種   教授
論文種別 原著
言語種別 英語
査読の有無 査読あり
表題 Fluorometric evaluation of CYP3A4 expression using improved transgenic HepaRG cells carrying a dual-colour reporter for CYP3A4 and CYP3A7.
掲載誌名 正式名:Scientific Reports
掲載区分国外
巻・号・頁 7(1),pp.2874
著者・共著者 Ueyama, T., Tsuji, S., Sugiyama, T., and Tada, M
担当区分 最終著者,責任著者
発行年月 2017/04
概要 Primary human hepatocytes are necessary to evaluate cytotoxicity, drug metabolism, and drug–drug
interactions for candidate compounds in early-phase drug discovery and development. However,
these analyses are often hampered by limited resources and functional or genetic variation among
lots. HepaRG human hepatocellular carcinoma cells can differentiate into mature hepatocyte-like cells (HepLCs) that possess similar metabolic activity to human hepatocytes. We previously established transgenic HepaRG cells carrying a dual reporter that express red fluorescent protein (RFP) under thetranscriptional regulation of CYP3A7 in the hepatoblast-like cell state and enhanced green fluorescent protein (EGFP) under the transcriptional regulation of CYP3A4 following HepLC differentiation. In this study, we successfully isolated a subclone of transgenic CYP3A4G/7R HepaRG cells with an improved HepLC differentiation potency. Midazolam metabolism by CYP3A4 in these HepLCs was comparable to that in wild-type HepLCs. The EGFP fluorescence intensity was greatly induced by rifampicin (RIF) treatment. There was a strong correlation between fluorometric and metabolic analyses. The fold change in EGFP-positive cells was comparable to those in the CYP3A4 mRNA level and luminescence of proluciferin metabolites. RIF treatment and cell proliferation increased the RFP-positive cell number. Thus, CYP3A4G/7R HepLCs provide a real-time, multiwell-based system to co-evaluate CYP3A4 induction and hepatic regeneration.