Inhibition of lysosome degradation on autophagosome formation and responses to GMI, an immunomodulatory protein from Ganoderma microsporum
Identifieur interne : 001453 ( Main/Exploration ); précédent : 001452; suivant : 001454Inhibition of lysosome degradation on autophagosome formation and responses to GMI, an immunomodulatory protein from Ganoderma microsporum
Auteurs : I-Lun Hsin [Taïwan] ; Gwo-Tarng Sheu [Taïwan] ; Ming-Shiou Jan [Taïwan] ; Hai-Lun Sun [Taïwan] ; Tzu-Chin Wu [Taïwan] ; Ling-Yen Chiu [Taïwan] ; Ko-Huang Lue [Taïwan] ; Jiunn-Liang Ko [Taïwan]Source :
- British journal of pharmacology [ 0007-1188 ] ; 2012.
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Abstract
BACKGROUND AND PURPOSE Autophagic cell death is considered a self-destructive process that results from large amounts of autophagic flux. In our previous study, GMI, a recombinant fungal immunomodulatory protein cloned from Ganoderma microsporum, induced autophagic cell death in lung cancer cells. The aim of this study was to examine the role of autophagosome accumulation in GMI-mediated cell death. EXPERIMENTAL APPROACH Western blot analysis, flow cytometry and confocal microscopy were used to evaluate the effects of different treatments, including silencing of ATP6V0A1 by use of short hairpin RNAi, on GMI-mediated cell death, lung cancer cell viability and autophagosome accumulation in vitro. KEY RESULTS Lysosome inhibitors bafilomycin-A1 and chloroquine increased GMI-mediated autophagic cell death. GMI and bafilomycin-A1 co-treatment induced the accumulation of large amounts of autophagosomes, but did not significantly induce apoptosis. GMI elicited autophagy through the PKB (Akt)/mammalian target of rapamycin signalling pathway. Silencing of ATP6V0A1, one subunit of vesicular H+-ATPases (V-ATPases) that mediates lysosome acidification, spontaneously induced autophagosome accumulation, but did not affect lysosome acidity. GMI-mediated autophagosome accumulation and cytotoxicity was increased in shATP6V0A1 lung cancer cells. Furthermore, ATP6V0A1 silencing decreased autophagosome and lysosome fusion in GMI-treated CaLu-1/GFP-LC3 lung cancer cells.
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i1="02"><s1>Institute of Medicine, Chung Shan Medical University</s1><s2>Taichung</s2><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation></author><author><name sortKey="Sheu, Gwo Tarng" sort="Sheu, Gwo Tarng" uniqKey="Sheu G" first="Gwo-Tarng" last="Sheu">Gwo-Tarng Sheu</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Medical and Molecular Toxicology, Chung Shan Medical University</s1><s2>Taichung</s2><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Institute of Medicine, Chung Shan Medical University</s1><s2>Taichung</s2><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation></author><author><name sortKey="Jan, Ming Shiou" sort="Jan, Ming Shiou" uniqKey="Jan M" first="Ming-Shiou" last="Jan">Ming-Shiou Jan</name><affiliation wicri:level="1"><inist:fA14 i1="06"><s1>Institute of Microbiology and Immunology, Chung Shan Medical University</s1><s2>Taichung</s2><s3>TWN</s3><sZ>3 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation></author><author><name sortKey="Sun, Hai Lun" sort="Sun, Hai Lun" uniqKey="Sun H" first="Hai-Lun" last="Sun">Hai-Lun Sun</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Division of Allergy, Department of Pediatrics, Chung Shan Medical University Hospital</s1><s2>Taichung</s2><s3>TWN</s3><sZ>4 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>School of Medicine, Chung Shall Medical University</s1><s2>Taichung</s2><s3>TWN</s3><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation></author><author><name sortKey="Wu, Tzu Chin" sort="Wu, Tzu Chin" uniqKey="Wu T" first="Tzu-Chin" last="Wu">Tzu-Chin Wu</name><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>School of Medicine, Chung Shall Medical University</s1><s2>Taichung</s2><s3>TWN</s3><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Division of Chest Medicine, Department of Internal Medicine, Chung Shan Medical University Hospital</s1><s2>Taichung</s2><s3>TWN</s3><sZ>5 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation></author><author><name sortKey="Chiu, Ling Yen" sort="Chiu, Ling Yen" uniqKey="Chiu L" first="Ling-Yen" last="Chiu">Ling-Yen Chiu</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Medical and Molecular Toxicology, Chung Shan Medical University</s1><s2>Taichung</s2><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Institute of Medicine, Chung Shan Medical University</s1><s2>Taichung</s2><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation></author><author><name sortKey="Lue, Ko Huang" sort="Lue, Ko Huang" uniqKey="Lue K" first="Ko-Huang" last="Lue">Ko-Huang Lue</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Institute of Medicine, Chung Shan Medical University</s1><s2>Taichung</s2><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Division of Allergy, Department of Pediatrics, Chung Shan Medical University Hospital</s1><s2>Taichung</s2><s3>TWN</s3><sZ>4 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>School of Medicine, Chung Shall Medical University</s1><s2>Taichung</s2><s3>TWN</s3><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation></author><author><name sortKey="Ko, Jiunn Liang" sort="Ko, Jiunn Liang" uniqKey="Ko J" first="Jiunn-Liang" last="Ko">Jiunn-Liang Ko</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Institute of Medical and Molecular Toxicology, Chung Shan Medical University</s1><s2>Taichung</s2><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Institute of Medicine, Chung Shan Medical University</s1><s2>Taichung</s2><s3>TWN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Division of Allergy, Department of Pediatrics, Chung Shan Medical University Hospital</s1><s2>Taichung</s2><s3>TWN</s3><sZ>4 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taichung</wicri:noRegion></affiliation></author></analytic><series><title level="j" type="main">British journal of pharmacology</title><title level="j" type="abbreviated">Br. j. pharmacol.</title><idno type="ISSN">0007-1188</idno><imprint><date when="2012">2012</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">British journal of pharmacology</title><title level="j" type="abbreviated">Br. j. pharmacol.</title><idno type="ISSN">0007-1188</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Adenosinetriphosphatase</term><term>Cell death</term><term>Ganoderma</term><term>Immunomodulator</term><term>Lysosome</term><term>Mammalian target of rapamycin</term><term>Microsporum</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Lysosome</term><term>Immunomodulateur</term><term>Protéine mTOR</term><term>Ganoderma</term><term>Microsporum</term><term>Mort cellulaire</term><term>Adenosinetriphosphatase</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">BACKGROUND AND PURPOSE Autophagic cell death is considered a self-destructive process that results from large amounts of autophagic flux. In our previous study, GMI, a recombinant fungal immunomodulatory protein cloned from Ganoderma microsporum, induced autophagic cell death in lung cancer cells. The aim of this study was to examine the role of autophagosome accumulation in GMI-mediated cell death. EXPERIMENTAL APPROACH Western blot analysis, flow cytometry and confocal microscopy were used to evaluate the effects of different treatments, including silencing of ATP6V0A1 by use of short hairpin RNAi, on GMI-mediated cell death, lung cancer cell viability and autophagosome accumulation in vitro. KEY RESULTS Lysosome inhibitors bafilomycin-A1 and chloroquine increased GMI-mediated autophagic cell death. GMI and bafilomycin-A1 co-treatment induced the accumulation of large amounts of autophagosomes, but did not significantly induce apoptosis. GMI elicited autophagy through the PKB (Akt)/mammalian target of rapamycin signalling pathway. Silencing of ATP6V0A1, one subunit of vesicular H<sup>+</sup>-ATPases (V-ATPases) that mediates lysosome acidification, spontaneously induced autophagosome accumulation, but did not affect lysosome acidity. GMI-mediated autophagosome accumulation and cytotoxicity was increased in shATP6V0A1 lung cancer cells. Furthermore, ATP6V0A1 silencing decreased autophagosome and lysosome fusion in GMI-treated CaLu-1/GFP-LC3 lung cancer cells.</div></front></TEI><affiliations><list><country><li>Taïwan</li></country></list><tree><country name="Taïwan"><noRegion><name sortKey="Hsin, I Lun" sort="Hsin, I Lun" uniqKey="Hsin I" first="I-Lun" last="Hsin">I-Lun Hsin</name></noRegion><name sortKey="Chiu, Ling Yen" sort="Chiu, Ling Yen" uniqKey="Chiu L" first="Ling-Yen" last="Chiu">Ling-Yen Chiu</name><name sortKey="Chiu, Ling Yen" sort="Chiu, Ling Yen" uniqKey="Chiu L" first="Ling-Yen" last="Chiu">Ling-Yen Chiu</name><name sortKey="Hsin, I Lun" sort="Hsin, I Lun" uniqKey="Hsin I" first="I-Lun" last="Hsin">I-Lun Hsin</name><name sortKey="Jan, Ming Shiou" sort="Jan, Ming Shiou" uniqKey="Jan M" first="Ming-Shiou" last="Jan">Ming-Shiou Jan</name><name sortKey="Ko, Jiunn Liang" sort="Ko, Jiunn Liang" uniqKey="Ko J" first="Jiunn-Liang" last="Ko">Jiunn-Liang Ko</name><name sortKey="Ko, Jiunn Liang" sort="Ko, Jiunn Liang" uniqKey="Ko J" first="Jiunn-Liang" last="Ko">Jiunn-Liang Ko</name><name sortKey="Ko, Jiunn Liang" sort="Ko, Jiunn Liang" uniqKey="Ko J" first="Jiunn-Liang" last="Ko">Jiunn-Liang Ko</name><name sortKey="Lue, Ko Huang" sort="Lue, Ko Huang" uniqKey="Lue K" first="Ko-Huang" last="Lue">Ko-Huang Lue</name><name sortKey="Lue, Ko Huang" sort="Lue, Ko Huang" uniqKey="Lue K" first="Ko-Huang" last="Lue">Ko-Huang Lue</name><name sortKey="Lue, Ko Huang" sort="Lue, Ko Huang" uniqKey="Lue K" first="Ko-Huang" last="Lue">Ko-Huang Lue</name><name sortKey="Sheu, Gwo Tarng" sort="Sheu, Gwo Tarng" uniqKey="Sheu G" first="Gwo-Tarng" last="Sheu">Gwo-Tarng Sheu</name><name sortKey="Sheu, Gwo Tarng" sort="Sheu, Gwo Tarng" uniqKey="Sheu G" first="Gwo-Tarng" last="Sheu">Gwo-Tarng Sheu</name><name sortKey="Sun, Hai Lun" sort="Sun, Hai Lun" uniqKey="Sun H" first="Hai-Lun" last="Sun">Hai-Lun Sun</name><name sortKey="Sun, Hai Lun" sort="Sun, Hai Lun" uniqKey="Sun H" first="Hai-Lun" last="Sun">Hai-Lun Sun</name><name sortKey="Wu, Tzu Chin" sort="Wu, Tzu Chin" uniqKey="Wu T" first="Tzu-Chin" last="Wu">Tzu-Chin Wu</name><name sortKey="Wu, Tzu Chin" sort="Wu, Tzu Chin" uniqKey="Wu T" first="Tzu-Chin" last="Wu">Tzu-Chin Wu</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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