The effect of paclitaxel on 2-aminofluorene-DNA adducts formation and arylamine N-acetyltransferase activity and gene expression in human lung tumor cells (A549).
In this study, paclitaxel was used to determine inhibition of arylamine N-acetyltransferase (NAT) activity, gene expression and 2-aminofluorene-DNA adduct formation in a human lung tumor cell line (A549). The activity of NAT was measured by HPLC assaying for the amounts of N-acetyl-2-aminofluorene (2-AAF) and remaining 2-aminofluorene (2-AF). Human lung tumor cell cytosols and intact cells were used for examining NAT activity and carcinogen-DNA adduct formation. The results demonstrated that NAT activity, gene expression (NAT1 mRNA) and 2-AF-DNA adduct formation in human lung tumor cells were inhibited and decreased by paclitaxel in a dose-dependent manner. The effects of paclitaxel on the values of the apparent Km and Vmax of NAT from human lung tumor cells were also determined in both examined systems. The result also indicated that paclitaxel decreased the apparent values of Km and Vmax from human lung tumor cells in both cytosol and intact cells. Thus, paclitaxel is an uncompetitive inhibitor to NAT enzyme.
Hsia
T C
TC
Department of Internal Medicine, China Medical College Hospital, No 2, Yuh-Der Road, Taichung 404, Taiwan, ROC.
Chung
J G
JG
Lu
H F
HF
Ho
H C
HC
Yang
C C
CC
Lu
K H
KH
Hung
C F
CF
D000107:Acetylation; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D001191:Arylamine N-Acetyltransferase / Q000235:genetics / Q000378:metabolism*; D002273:Carcinogens / Q000737:chemistry*; D003600:Cytosol / Q000201:enzymology / Q000378:metabolism; D018736:DNA Adducts / Q000187:drug effects*; D003864:Depression, Chemical; D004305:Dose-Response Relationship, Drug; D005449:Fluorenes / Q000737:chemistry*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D007700:Kinetics; D008175:Lung Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D017239:Paclitaxel / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D000972:Antineoplastic Agents, Phytogenic; D002273:Carcinogens; D018736:DNA Adducts; D005449:Fluorenes; C012177:2-aminofluorene; D001191:Arylamine N-Acetyltransferase; D017239:Paclitaxel