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Generation of hydrogen peroxide precedes loss of mitochondrial membrane potential during DNA alkylation‐induced apoptosis
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Pulsed field gel electrophoresis showed that the initiation time of DNA breakage induced by the DNA alkylating agent duocarmycin A, which is not a redox‐cycling agent, was almost the same in the human leukemia cell line HL‐60 and its H2O2‐resistant clone HP100. Catalase activity of HP100 cells was much higher than that of HL‐60 cells. Duocarmycin A‐mediated DNA ladder formation in HP100 cells was delayed compared with that in HL‐60 cells, suggesting the involvement of H2O2 in duocarmycin A‐induced apoptosis. Flow cytometry demonstrated that peroxide formation preceded loss of mitochondrial membrane potential (ΔΨm) in cells treated with duocarmycin A. Then, caspase‐3 was activated, followed by DNA ladder formation. These findings suggest that DNA damage by duocarmycin A induces H2O2 generation, which causes ΔΨm loss and subsequently caspase‐3 activation, resulting in apoptosis.
Title: Generation of hydrogen peroxide precedes loss of mitochondrial membrane potential during DNA alkylation‐induced apoptosis
Description:
Pulsed field gel electrophoresis showed that the initiation time of DNA breakage induced by the DNA alkylating agent duocarmycin A, which is not a redox‐cycling agent, was almost the same in the human leukemia cell line HL‐60 and its H2O2‐resistant clone HP100.
Catalase activity of HP100 cells was much higher than that of HL‐60 cells.
Duocarmycin A‐mediated DNA ladder formation in HP100 cells was delayed compared with that in HL‐60 cells, suggesting the involvement of H2O2 in duocarmycin A‐induced apoptosis.
Flow cytometry demonstrated that peroxide formation preceded loss of mitochondrial membrane potential (ΔΨm) in cells treated with duocarmycin A.
Then, caspase‐3 was activated, followed by DNA ladder formation.
These findings suggest that DNA damage by duocarmycin A induces H2O2 generation, which causes ΔΨm loss and subsequently caspase‐3 activation, resulting in apoptosis.
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