Exp Mol Med.
2000 Mar;32(1):42-46.
Regulation of bcl-2 family in hydrogen peroxide-induced apoptosis in human leukemia HL-60 cells
- Affiliations
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- 1Department of Pediatrics, Korea University, Seoul.
Abstract
- Numerous types of cells have been shown to undergo apoptosis when exposed to oxidant agent such as hydrogen peroxide. In order to understand the functional relationship between the anti- and pro-apoptotic regulatory proteins in the cells under oxidant stress, we have studied the level of expression of apoptosis regulatory proteins, bcl-2 and bax, in human leukemia HL-60 cells. The exposure of HL-60 cells to different concentrations of H2O2 for 6 h resulted in a typical apoptosis of the cells as characterized by flow cytometry, cell cycle analysis, and DNA fragmantation. There was a block in G1 to S transition and apoptotic cells were mainly derived from S and G2 cells. Kinetic study demonstrated that the levels of both bcl-2-mRNA and -protein expression were decreased with the progression of cellular apoptosis whereas the level of bax-mRNA was unchanged but the expressed bax-protein was not detectable. Cycloheximide, a nonspecific translation inhibitor, did not prevent the hydrogen peroxide-mediated apoptosis in HL-60 cells. These results suggest that the regulation of bcl-2, but not of bax are important factor in the oxidative stress-induced apoptosis in HL-60 cells.
Keyword
MeSH Terms
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Apoptosis/drug effects*
Blotting, Western
Cycloheximide/pharmacology
DNA Fragmentation
DNA, Neoplasm/metabolism
DNA, Neoplasm/genetics
DNA, Neoplasm/drug effects
Dose-Response Relationship, Drug
Flow Cytometry
Gene Expression Regulation, Neoplastic/drug effects
HL-60 Cells
Human
Hydrogen Peroxide/pharmacology*
Oxidants/pharmacology*
Protein Synthesis Inhibitors/pharmacology
Proto-Oncogene Proteins/metabolism
Proto-Oncogene Proteins/genetics
Proto-Oncogene Proteins c-bcl-2/metabolism
Proto-Oncogene Proteins c-bcl-2/genetics*
RNA, Messenger/metabolism
RNA, Messenger/genetics
RNA, Messenger/drug effects
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