Establishment of a human hepatoma cell line, HLE/2E1, suitable for detection of P450 2E1-related cytotoxicity

Isao Nozaki, Toshiya Tsuji, Masakiyo Sakaguchi, Yusuke Inoue, Ryuji Hirai, Akio Andou, Masahiro Miyazaki, Nobuyoshi Shimizu, Masayoshi Namba

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

By transfection of an expression vector of human cytochrome P450 2E1 (CYP2E1) into a human hepatoma cell line (HLE), a new cell line (HLE/2E1) that stably expresses activity of CYP2E1 has been established. The HLE/2E1 cell line expressed a higher level of CYP2E1 messenger ribonucleic acid than did the mother HLE cell line. CYP2E1 enzyme activity determined by a p-nitrophenol oxidation assay was also higher in HLE/2E1 cells than in HLE cells. In addition, the enzyme activity of the HLE/2E1 cells was increased by ethanol treatment. Exposure to acetaminophen (APAP) or buthionine sulfoximine (BSO) caused a greater decrease in viability of the HLE/2E1 cells than that of the HLE cells, as determined by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. The cytotoxicity of APAP or BSO to HLE/2E1 cells was inhibited by the addition of ethanol or vitamin E. However, the cytotoxicity of both APAP and BSO was enhanced by 24-h preincubation of HLE/2E1 cells with ethanol. These results show that this cell line provides a useful model for studying catalytic properties of CYP2E1 and cytotoxic mechanisms of chemicals metabolized by CYP2E1.

Original languageEnglish
Pages (from-to)566-570
Number of pages5
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume36
Issue number9
DOIs
Publication statusPublished - Jan 1 2000

Keywords

  • Acetaminophen
  • Buthionine sulfoximine
  • CYP2E1
  • Ethanol
  • Vitamin E

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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