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

I. Nozaki, T. Tsuji, Masakiyo Sakaguchi, Y. Inoue, R. Hirai, A. Andou, M. Miyazaki, N. Shimizu, M. 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 - 2000

Fingerprint

Cytochrome P-450 CYP2E1
Cytotoxicity
Hepatocellular Carcinoma
Cells
Cell Line
Acetaminophen
Buthionine Sulfoximine
Ethanol
Enzyme activity
Assays
Vitamin E
RNA
Oxidation
Cytochrome P-450 Enzyme System
Transfection

Keywords

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

ASJC Scopus subject areas

  • Developmental Biology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Establishment of a human hepatoma cell line, HLE/2E1, suitable for detection of P450 2E1-related cytotoxicity. / Nozaki, I.; Tsuji, T.; Sakaguchi, Masakiyo; Inoue, Y.; Hirai, R.; Andou, A.; Miyazaki, M.; Shimizu, N.; Namba, M.

In: In Vitro Cellular and Developmental Biology - Animal, Vol. 36, No. 9, 2000, p. 566-570.

Research output: Contribution to journalArticle

Nozaki, I. ; Tsuji, T. ; Sakaguchi, Masakiyo ; Inoue, Y. ; Hirai, R. ; Andou, A. ; Miyazaki, M. ; Shimizu, N. ; Namba, M. / Establishment of a human hepatoma cell line, HLE/2E1, suitable for detection of P450 2E1-related cytotoxicity. In: In Vitro Cellular and Developmental Biology - Animal. 2000 ; Vol. 36, No. 9. pp. 566-570.
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AU - Inoue, Y.

AU - Hirai, R.

AU - Andou, A.

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