Wogonin inhibits N-acetyltransferase activity and gene expression in human leukemia HL-60 cells.
It is well documented that arylamine carcinogens are N-acetylated by cytosolic N-acetyltransferase (NAT) enzyme. NAT plays an important role in the metabolizing of those arylamine compounds. 2-Aminofluorene (AF) is an arylamine carcinogen which has been demonstrated to induce carcinogenesis in laboratory animals. Our previous study has shown that a human promyelocytic leukemia cell line, HL-60, displays NAT activity. The purpose of the present study was to determine whether or not wogonin could affect the N-acetylation of AF in HL-60. N-acetylated and non-N-acetylated AF were determined by using high performance liquid chromatography. Wogonin displayed a dose-dependent inhibition of NAT activity in cytosols and intact cells. Wogonin also decreased AF-DNA adduct formation in these cells. The effects of wogonin on the NAT enzymes levels were also examined by Western blotting and flow cytometry and the changes of NAT gene expression were examined by polymerase chain reaction (PCR) and cDNA microarray. The results demonstrated that wogonin inhibited NAT1 mRNA gene expression and the level of NAT enzyme in HL-60 cells. This is the first demonstration that wogonin affects human leukemia cells' NAT activity in vitro.
Yu
Chun-Shu
CS
Center of General Education, China Medical University, Taichung, Taiwan, ROC.
Yu
Fu-Shun
FS
Chuang
Yao-Chen
YC
Lu
Hsu-Feng
HF
Lin
Shuw-Yun
SY
Chiu
Tsan-Hung
TH
Chung
Jing-Gung
JG
D015073:2-Acetylaminofluorene / Q000031:analogs & derivatives* / Q000378:metabolism; D000107:Acetylation / Q000187:drug effects; D000123:Acetyltransferases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D018736:DNA Adducts / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D044950:Flavanones / Q000494:pharmacology*; D005449:Fluorenes / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D018922:HL-60 Cells; D006801:Humans; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics
C052668:2-aminofluorene-DNA complex; D018736:DNA Adducts; D004791:Enzyme Inhibitors; D044950:Flavanones; D005449:Fluorenes; D012333:RNA, Messenger; C012177:2-aminofluorene; D015073:2-Acetylaminofluorene; D000123:Acetyltransferases; C085514:wogonin