Ketoprofen-inhibited N-acetyltransferase activity and gene expression in human colon tumor cells.
The activation of ketoprofen, which inhibits the outgrowth of azoxymethane-induced aberrant crypt foci in the rat colon, on the inhibition of arylamine N-acetyltransferase (NAT) activity (N-acetylation of substrates), gene expression (mRNA NAT) and 2-aminofluorene (AF)-DNA adduct formation was studied in a human colon tumor (adenocarcinoma) cell line (colo 205). Cellular cytosols (9000 xg supernatant) and intact colon tumor cells were used. The NAT activity in colo 205 cells was inhibited by ketoprofen in a dose- and time -dependent manner in both examined systems. The data also indicated that ketoprofen decreased the apparent value of V(max) of NAT enzymes, being a competitive inhibitor of NAT enzymes. The AF-DNA adduct formation in colo 205 cells was also decreased by ketoprofen. Based on the results from PCR, it was shown that ketoprofen affected mRNA NAT expression in human colon colo 205 cells. The cells were stained with anti-NAT antibody, then analyzed by flow cytometry. The results showed that ketoprofen decreased the percentage of cells stained by anti-NAT. This report is the first to demonstrate that ketoprofen inhibits human colon tumor cell NAT activity, gene expression and DNA adduct formation.
Cheng
Kwork-Chu
KC
Department of Surgery, Jen-Ai Hospital, 483, Tong-Rong Road, Tali, Taichung, Taiwan, ROC.
Li
Yu-Ching
YC
Yu
Chun-Su
CS
Yu
Fu-Shun
FS
Lee
Jau-Hong
JH
Lin
Meng-Liang
ML
Yang
Jai-Sing
JS
Chung
Jing-Gung
JG
D015073:2-Acetylaminofluorene / Q000031:analogs & derivatives / Q000378:metabolism; D010129:4-Aminobenzoic Acid / Q000378:metabolism; D000107:Acetylation / Q000187:drug effects; D001191:Arylamine N-Acetyltransferase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000188:drug therapy* / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D018736:DNA Adducts / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D005449:Fluorenes; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007660:Ketoprofen / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D062366:para-Aminobenzoates
C052668:2-aminofluorene-DNA complex; D018736:DNA Adducts; D005449:Fluorenes; D012333:RNA, Messenger; D062366:para-Aminobenzoates; C023730:Acedoben; D007660:Ketoprofen; D015073:2-Acetylaminofluorene; D001191:Arylamine N-Acetyltransferase; D010129:4-Aminobenzoic Acid