Involvement of N-acetyltransferase human in the cytotoxic activity of 5-fluorouracil.
N-acetyltransferase human (NATH) participates in a posttranslational modification of the proteins and has been reported to play a role in apoptosis. In this study, the involvement of NATH in the cytotoxic action of 5-fluorouracil (5-FU) in human squamous cell carcinoma HEp-2 cells was examined. We found that 5-FU decreased NATH expression in a dose-dependent and time-dependent manner. No change was observed after treatment with bleomycin, nedaplatin, mitomycin C, or methotrexate. Interestingly, knockdown of NATH by small interfering RNA resulted in the downregulation of thymidylate synthase mRNA expression and induced apoptosis. Conversely, NATH overexpression facilitated cell proliferation independent of the presence of 5-FU. The effect of NATH knockdown on the expression of proteins in HEp-2 cells was examined using two-dimensional gel electrophoresis and mass spectrometry. Profilin 1, CutA, ras-related nuclear protein, annexin A5, enolase 1, and elongation factor 1 alpha 1 were found to be upregulated and 14-3-3eta, tublin, nuclear auto antigenic sperm protein, heat shock protein 70, and heat shock protein 90 were downregulated by knockdown of NATH. The results of this study suggest that NATH plays an important role in the cytotoxic activity of 5-FU.
Takubo
Kazuko
K
Division of Molecular and Genetic Medicine, Department of Genetic Medicine and Regenerative Therapeutics, Graduate School of Medicine, Tottori University, Nishimachi 36-1, Yonago 683-8504, Japan. kazuko@med.tottori-u.ac.jp
Tsuchiya
Hiroyuki
H
Kurimasa
Akihiro
A
Arnesen
Thomas
T
Ryoke
Kazuo
K
Shiota
Goshi
G
D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001191:Arylamine N-Acetyltransferase / Q000235:genetics / Q000378:metabolism*; D053453:Caspase 9 / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D015536:Down-Regulation / Q000187:drug effects; D005472:Fluorouracil / Q000494:pharmacology* / Q000633:toxicity; D015870:Gene Expression / Q000187:drug effects / Q000235:genetics; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects; D011506:Proteins / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D013940:Thymidylate Synthase / Q000235:genetics; D014162:Transfection; D015854:Up-Regulation / Q000187:drug effects
D000970:Antineoplastic Agents; D007527:Isoenzymes; D011506:Proteins; D034741:RNA, Small Interfering; D013940:Thymidylate Synthase; D001191:Arylamine N-Acetyltransferase; C089384:N-acetyltransferase 1; D011065:Poly(ADP-ribose) Polymerases; D053453:Caspase 9; D005472:Fluorouracil