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 language | English |
|---|---|
| Pages (from-to) | 566-570 |
| Number of pages | 5 |
| Journal | In Vitro Cellular and Developmental Biology - Animal |
| Volume | 36 |
| Issue number | 9 |
| DOIs | |
| Publication status | Published - 2000 |
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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 journal › Article
}
TY - JOUR
T1 - Establishment of a human hepatoma cell line, HLE/2E1, suitable for detection of P450 2E1-related cytotoxicity
AU - Nozaki, I.
AU - Tsuji, T.
AU - Sakaguchi, Masakiyo
AU - Inoue, Y.
AU - Hirai, R.
AU - Andou, A.
AU - Miyazaki, M.
AU - Shimizu, N.
AU - Namba, M.
PY - 2000
Y1 - 2000
N2 - 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.
AB - 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.
KW - Acetaminophen
KW - Buthionine sulfoximine
KW - CYP2E1
KW - Ethanol
KW - Vitamin E
UR - http://www.scopus.com/inward/record.url?scp=0034496123&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034496123&partnerID=8YFLogxK
U2 - 10.1290/1071-2690(2000)036<0566:EOAHHC>2.0.CO;2
DO - 10.1290/1071-2690(2000)036<0566:EOAHHC>2.0.CO;2
M3 - Article
C2 - 11212141
AN - SCOPUS:0034496123
VL - 36
SP - 566
EP - 570
JO - In Vitro Cellular and Developmental Biology - Plant
JF - In Vitro Cellular and Developmental Biology - Plant
SN - 1071-2690
IS - 9
ER -