| Vacuolar H+-ATPase inhibitor induces apoptosis via lysosomal dysfunction in the human gastric cancer cell line MKN-1. | |
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MedLine Citation:
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PMID: 14561721 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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We investigated the mechanism of apoptosis induced by bafilomycin A(1), an inhibitor of vacuolar H(+)-ATPase. Bafilomycin A(1) significantly inhibited the growth of MKN-1 human gastric cancer cells. Bafilomycin A(1) induced apoptosis as demonstrated by DNA ladder formation and the TUNEL method. We designed a flow cytometric assay to detect the alteration in lysosomal pH using a fluorescent probe, fluorescein isothiocyanate-conjugated dextran. This assay revealed that bafilomycin A(1) dramatically increased lysosomal pH. However, bafilomycin A(1) induced neither significant decrease in mitochondrial transmembrane potential nor the release of mitochondrial cytochrome c into the cytoplasm. Western blotting showed that cathepsin D, but not cathepsin L, was released into the cytoplasm. The activity of caspase-3 was significantly increased by bafilomycin A(1). However, cathepsin D did not directly cleave procaspase-3. These findings suggest that bafilomycin A(1)-induced apoptosis in MKN-1 cells is mediated by other proteases released after lysosomal dysfunction followed by activation of caspase-3 in a cytochrome c-independent manner. The present study showed that flow cytometric analysis of lysosomal pH can be useful to evaluate lysosomal protease-mediated apoptosis. |
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Authors:
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Shigehito Nakashima; Yusuke Hiraku; Saeko Tada-Oikawa; Terutoshi Hishita; Esteban C Gabazza; Shigenori Tamaki; Ichiro Imoto; Yukihiko Adachi; Shosuke Kawanishi |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Journal of biochemistry Volume: 134 ISSN: 0021-924X ISO Abbreviation: J. Biochem. Publication Date: 2003 Sep |
Date Detail:
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Created Date: 2003-10-16 Completed Date: 2004-09-14 Revised Date: 2007-12-19 |
Medline Journal Info:
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Nlm Unique ID: 0376600 Medline TA: J Biochem Country: Japan |
Other Details:
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Languages: eng Pagination: 359-64 Citation Subset: IM |
Affiliation:
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Third Department of Internal Medicine, Mie University School of Medicine, Tsu, Mie, 514-8507. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Apoptosis
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drug effects* Caspase 3 Caspases / metabolism Cathepsin D / metabolism Cell Line, Tumor Enzyme Inhibitors / pharmacology Flow Cytometry Humans Hydrogen-Ion Concentration In Situ Nick-End Labeling Lysosomes / metabolism Macrolides / pharmacology* Proton-Translocating ATPases / antagonists & inhibitors |
| Chemical | |
Reg. No./Substance:
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0/Enzyme Inhibitors; 0/Macrolides; 88899-55-2/bafilomycin A1; EC 3.4.22.-/CASP3 protein, human; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspases; EC 3.4.23.5/Cathepsin D; EC 3.6.3.14/Proton-Translocating ATPases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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