PCR and flow cytometric analysis of paclitaxel-inhibited arylamine N-acetyltransferase activity and gene expression in human osteogenic sarcoma cells (U-2 OS).

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.

• Authors: Ko-Hsiu Lu, Der-Yean Wang, Ko-Huang Lue, Yie-Ming Hsiao, Ming-Chih Chou, Yi-Shuan Chen, Jing-Gung Chung
• Journal: Anticancer research (Anticancer Res) 2004 Jan-Feb Vol. 24 Issue 1 Pg. 83-90 ISSN: 0250-7005 [Print] Greece
• PMID: 15015580 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• 2-aminofluorene-DNA complex
• Antineoplastic Agents, Phytogenic
• DNA Adducts
• Fluorenes
• RNA, Messenger
• para-Aminobenzoates
• Acedoben
• 2-Acetylaminofluorene
• Arylamine N-Acetyltransferase
• Paclitaxel
• 4-Aminobenzoic Acid

Topics

• 2-Acetylaminofluorene (analogs & derivatives, metabolism)
• 4-Aminobenzoic Acid (metabolism)
• Adolescent
• Antineoplastic Agents, Phytogenic (pharmacology)
• Arylamine N-Acetyltransferase (antagonists & inhibitors, biosynthesis, genetics, metabolism)
• Bone Neoplasms (drug therapy, enzymology, genetics)
• Cell Line, Tumor
• Cytosol (enzymology)
• DNA Adducts (antagonists & inhibitors, biosynthesis)
• Dose-Response Relationship, Drug
• Female
• Flow Cytometry
• Fluorenes (antagonists & inhibitors)
• Gene Expression (drug effects)
• Humans
• Osteosarcoma (drug therapy, enzymology, genetics)
• Paclitaxel (pharmacology)
• Polymerase Chain Reaction
• RNA, Messenger (biosynthesis, genetics)
• para-Aminobenzoates