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
  • 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

hydrogen peroxide; apoptosis; HL-60 cell; Bcl-2; cell cycle

MeSH Terms

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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