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Specific inhibitors of vacuolar type H(+)-ATPases induce apoptotic cell death
T Nishihara1, S Akifusa, T Koseki
1Department of Oral Science, National Institute of Health, Tokyo, Japan.
Abstract:
Concanamycin A and bafilomycin A1 are known as strong inhibitors of the vacuolar type H(+)-ATPases in vitro. These inhibitors exhibited cytotoxic effects on twelve cell lines in cell viability assay. On the other hand, the F1F0-type H(+)-ATPase inhibitor oligomycin and the E1E2-type H(+)-ATPase inhibitor vanadate showed no cytotoxic effect. We show here that concanamycin A and bafilomycin A1 induce a significant increase in the proportion of fragmented DNA in agarose gel electrophoresis. Flow cytometric cell cycle analysis of WEHI 231 cells stimulated with concanamycin A revealed the increased percentage of apoptotic cells with hypodiploid DNA. These findings indicate that cell death induced by specific inhibitors of vacuolar type H(+)-ATPases occurs through apoptosis.
Insights
Specific vacuolar type H(+)-ATPase inhibitors, concanamycin A and bafilomycin A1, induce apoptosis. This cell death pathway was confirmed through DNA fragmentation and flow cytometry analysis in multiple cell lines.
Area of Science:
- Cell biology
- Biochemistry
- Molecular biology
Background:
- Vacuolar type H(+)-ATPases are crucial for cellular processes.
- Concanamycin A and bafilomycin A1 are known inhibitors of vacuolar H(+)-ATPases.
- The cytotoxic effects of these inhibitors on various cell lines warrant further investigation into the underlying cell death mechanisms.
Purpose of the Study:
- To investigate the mechanism of cell death induced by concanamycin A and bafilomycin A1.
- To determine if vacuolar type H(+)-ATPase inhibition leads to apoptosis.
- To differentiate the effects of vacuolar H(+)-ATPase inhibitors from other ATPase inhibitors.
Main Methods:
- Cell viability assays on twelve cell lines.
- Agarose gel electrophoresis to detect DNA fragmentation.
- Flow cytometric cell cycle analysis to assess apoptosis.
Main Results:
- Concanamycin A and bafilomycin A1 exhibited cytotoxic effects, unlike oligomycin and vanadate.
- Inhibitors of vacuolar type H(+)-ATPases significantly increased DNA fragmentation.
- Flow cytometry confirmed an increased percentage of apoptotic cells with hypodiploid DNA.
Conclusions:
- Cell death induced by specific vacuolar type H(+)-ATPase inhibitors occurs via apoptosis.
- Vacuolar type H(+)-ATPase inhibition is a potent inducer of programmed cell death.
- These findings highlight the critical role of vacuolar H(+)-ATPases in maintaining cell viability.