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Research ArticleExperimental Studies

PCR and Flow Cytometric Analysis of Paclitaxel-inhibited Arylamine N-Acetyltransferase Activity and Gene Expression in Human Osteogenic Sarcoma Cells (U-2 OS)

KO-HSIU LU, DER-YEAN WANG, KO-HUANG LUE, YIE-MING HSIAO, MING-CHIH CHOU, YI-SHUAN CHEN and JING-GUNG CHUNG
Anticancer Research January 2004, 24 (1) 83-90;
KO-HSIU LU
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DER-YEAN WANG
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KO-HUANG LUE
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YIE-MING HSIAO
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MING-CHIH CHOU
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YI-SHUAN CHEN
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JING-GUNG CHUNG
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Abstract

Human epidemiological studies suggest an association between N-acetyltransferase (NAT) activity and the incidence of bladder and colorectal cancers. In this study, paclitaxel was selected to examine the inhibition of arylamine NAT activity, gene expression and 2-aminofluorene-DNA adduct formation in a human osteogenic sarcoma cell line (U-2 OS). The activity of NAT was determined by high performance liquid chromatography (HPLC) assay for the amounts of acetylated 2-aminofluorene (AF) and p-aminobenzoic acid (PABA) and nonacetylated AF and PABA. Human osteogenic sarcoma cell cytosols and intact cells were used to examine the NAT activity, gene expression and AF-DNA adduct formation. The results demonstrated that NAT activity, percent of NAT in examined cells, gene expression (NAT1 mRNA) and AF-DNA adduct formation in human osteogenic sarcoma cells were inhibited and decreased by paclitaxel in a dose-dependent manner. The results also demonstrated that paclitaxel decreased the apparent values of Km and Vmax from intact human osteogenic sarcoma cells (U-2 OS). Thus, paclitaxel is an uncompetitive inhibitor of the NAT enzyme.

  • Received May 9, 2003.
  • Accepted December 2, 2003.
  • Copyright© 2004 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved
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Anticancer Research: 24 (1)
Anticancer Research
Vol. 24, Issue 1
January-February 2004
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PCR and Flow Cytometric Analysis of Paclitaxel-inhibited Arylamine N-Acetyltransferase Activity and Gene Expression in Human Osteogenic Sarcoma Cells (U-2 OS)
KO-HSIU LU, DER-YEAN WANG, KO-HUANG LUE, YIE-MING HSIAO, MING-CHIH CHOU, YI-SHUAN CHEN, JING-GUNG CHUNG
Anticancer Research Jan 2004, 24 (1) 83-90;

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PCR and Flow Cytometric Analysis of Paclitaxel-inhibited Arylamine N-Acetyltransferase Activity and Gene Expression in Human Osteogenic Sarcoma Cells (U-2 OS)
KO-HSIU LU, DER-YEAN WANG, KO-HUANG LUE, YIE-MING HSIAO, MING-CHIH CHOU, YI-SHUAN CHEN, JING-GUNG CHUNG
Anticancer Research Jan 2004, 24 (1) 83-90;
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