Apoptosis induced by hydrogen peroxide under serum deprivation and its inhibition by antisense c-jun in F-MEL cells.
Under serum deprivation F-MEL cells die by apoptosis. We previously showed that apoptosis induced by serum deprivation was suppressed by inhibition of c-jun expression using antisense c-jun transfected cell line, c-junAS. To elucidate the underlying mechanisms we examined the species which is responsible for apoptosis under serum deprivation. When catalase and N-acetyl-L-cysteine (NAC) were included in the medium, cell death under serum deprivation was effectively suppressed in F-MEL cells. Intracellular generation of hydrogen peroxide (H(2)O(2)) was also detected under serum deprivation in parental F-MEL cells, but it was suppressed in c-junAS (+) cells, in which antisense c-jun was expressed and c-Jun protein expression was inhibited as shown by Western blot. When H(2)O(2) was directly applied to F-MEL cells at 3 mM, apoptotic cell death was induced, whereas it was suppressed in c-junAS (+) cells. Induction of apoptosis by H(2)O(2) and its inhibition by antisense c-jun was confirmed by detection of internucleosomal fragmentation of DNA, TdT-mediated dUTP nick end labeling (TUNEL)-positive cells and morphological alteration of nuclei. These results indicate that apoptosis induced by serum deprivation in F-MEL cells is mediated by H(2)O(2) and c-jun expression is essential to apoptosis induced by H(2)O(2) in F-MEL cells.
Kim
Y H
YH
Research Center for Advanced Science and Technology, University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan. sarah@oxygen.rcast.u-tokyo.ac.jp
Takahashi
M
M
Suzuki
E
E
Niki
E
E
D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002374:Catalase / Q000494:pharmacology; D016895:Culture Media, Serum-Free; D053938:DNA Fragmentation / Q000187:drug effects; D016373:DNA, Antisense / Q000494:pharmacology*; D005434:Flow Cytometry; D005622:Friend murine leukemia virus; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D016758:Genes, jun / Q000235:genetics*; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D020287:In Situ Nick-End Labeling; D004915:Leukemia, Erythroblastic, Acute; D051379:Mice; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism; D014407:Tumor Cells, Cultured
D016895:Culture Media, Serum-Free; D016373:DNA, Antisense; D016755:Proto-Oncogene Proteins c-jun; D006861:Hydrogen Peroxide; D002374:Catalase; D000111:Acetylcysteine