Analysis of the intracellular regulatory mechanism of bcl-2 related apoptosis.

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1. Anti-apoptotic effect and the expression of p26 bcl-2 We have investigated the anti-apoptotic effect of bcl-2 proto-oncogene using bcl-2 transfectants (bcl-2 was introduced to CCRF-CEM,MBC-1, WEHI-164,2AD4) with various factor inducible apoptosis. The cells expressing a large amount of p26 bcl-2 showed higher viability when cell death was induced with H_2O_2, Dexamethasone and anti-lgM.Especially, bcl-2 has a stronger anti-apoptotic effect in the death induced with H_2O_2 than in the death induced with others. There was no significant correlation between the anti-apoptotic effect and the expression of p26 bcl-2 in the cell death induced with A23187, TNF-alpha. 2. Intarcellular events in the early phase of apoptosis and the expression of p26 bcl-2 The studies of intracellular reactive oxygen species and mitochondrial transmembrane potential of MBC-1 cells using fluorescence dye revealed no significant difference between bcl-2 transfectants and controls when cell death was induced with H_2O_2. Whereas p26 bcl-2 has significant anti-apoptotic effect in MBC-1 cells undergoing apoptosis with H_2O_2. When the transfectants of CEM-C7 cells were treated with H_2O_2, reactive oxygen species were markedly reduced but not correlate with the level of p26 bcl-2. Both intracellular reactive oxygen species and mitochondrial transmembrane potential showed no significant change in CEM,MBC-1 cells when cell death was induced with dexamethasone and anti-lgM respectively. These results suggest the anti-apoptotic effect of p26 bcl-2 has some direct relation to reactive oxygen species in early phase of apoptosis.