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Knockdown of Atg7 Induces Nuclear-LC3 Dependent Apoptosis and Augments Chemotherapy in Colorectal Cancer Cells

Identifieur interne : 000124 ( an2020/Analysis ); précédent : 000123; suivant : 000125

Knockdown of Atg7 Induces Nuclear-LC3 Dependent Apoptosis and Augments Chemotherapy in Colorectal Cancer Cells

Auteurs : Anna-Lena Scherr ; Adam Jassowicz ; Anna Pat ; Christin Elssner ; Lars Ismail ; Nathalie Schmitt ; Paula Hoffmeister ; Lasse Neukirch ; Georg Gdynia ; Benjamin Goeppert ; Henning Schulze-Bergkamen ; Dirk J Ger ; Bruno Christian Köhler

Source :

RBID : PMC:7038172

Abstract

Autophagy is a catabolic process that enables cells to degrade obsolete content and refuel energy depots. In colorectal cancer (CRC) autophagy has been shown to promote tumorigenesis through energy delivery in the condition of uncontrolled proliferation. With this study, we aimed at evaluating whether autophagy sustains CRC cell viability and if it impacts therapy resistance. Initially, a colorectal cancer tissue micro array, containing mucosa (n = 10), adenoma (n = 18) and adenocarcinoma (n = 49) spots, was stained for expression of essential autophagy proteins LC3b, Atg7, p62 and Beclin-1. Subsequently, central autophagy proteins were downregulated in CRC cells using siRNA technology. Viability assays, flow cytometry and immunoblotting were performed and three-dimensional cell culture was utilized to study autophagy in a tissue mimicking environment. In our study we found an upregulation of Atg7 in CRC. Furthermore, we identified Atg7 as crucial factor within the autophagy network for CRC cell viability. Its disruption induced cell death via triggering apoptosis and in combination with conventional chemotherapy it exerted synergistic effects in inducing CRC cell death. Cell death was strictly dependent on nuclear LC3b, since simultaneous knockdown of Atg7 and LC3b completely restored viability. This study unravels a novel cell death preventing function of Atg7 in interaction with LC3b, thereby unmasking a promising therapeutic target in CRC.


Url:
DOI: 10.3390/ijms21031099
PubMed: 32046105
PubMed Central: 7038172


Affiliations:


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PMC:7038172

Le document en format XML

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<title xml:lang="en" level="a" type="main">Knockdown of Atg7 Induces Nuclear-LC3 Dependent Apoptosis and Augments Chemotherapy in Colorectal Cancer Cells</title>
<author>
<name sortKey="Scherr, Anna Lena" sort="Scherr, Anna Lena" uniqKey="Scherr A" first="Anna-Lena" last="Scherr">Anna-Lena Scherr</name>
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<nlm:aff id="af1-ijms-21-01099">National Center for Tumor Diseases, University Hospital Heidelberg, Heidelberg 69120, Germany;
<email>anna-lena.scherr@nct-heidelberg.de</email>
(A.-L.S.);
<email>adam@jassowicz.com</email>
(A.J.);
<email>patoannaterezia@gmail.com</email>
(A.P.);
<email>Christin.Elssner@gmx.de</email>
(C.E.);
<email>lars.ismail1986@googlemail.com</email>
(L.I.);
<email>nathalie.schmitt@nct-heidelberg.de</email>
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(D.J.)</nlm:aff>
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<name sortKey="Jassowicz, Adam" sort="Jassowicz, Adam" uniqKey="Jassowicz A" first="Adam" last="Jassowicz">Adam Jassowicz</name>
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<nlm:aff id="af1-ijms-21-01099">National Center for Tumor Diseases, University Hospital Heidelberg, Heidelberg 69120, Germany;
<email>anna-lena.scherr@nct-heidelberg.de</email>
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<email>adam@jassowicz.com</email>
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(A.P.);
<email>Christin.Elssner@gmx.de</email>
(C.E.);
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(L.I.);
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(D.J.)</nlm:aff>
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<name sortKey="Pat, Anna" sort="Pat, Anna" uniqKey="Pat A" first="Anna" last="Pat">Anna Pat</name>
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<nlm:aff id="af1-ijms-21-01099">National Center for Tumor Diseases, University Hospital Heidelberg, Heidelberg 69120, Germany;
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<name sortKey="Elssner, Christin" sort="Elssner, Christin" uniqKey="Elssner C" first="Christin" last="Elssner">Christin Elssner</name>
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<name sortKey="Ismail, Lars" sort="Ismail, Lars" uniqKey="Ismail L" first="Lars" last="Ismail">Lars Ismail</name>
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(L.I.);
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(N.S.);
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(P.H.);
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(D.J.)</nlm:aff>
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<name sortKey="Schmitt, Nathalie" sort="Schmitt, Nathalie" uniqKey="Schmitt N" first="Nathalie" last="Schmitt">Nathalie Schmitt</name>
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<name sortKey="Hoffmeister, Paula" sort="Hoffmeister, Paula" uniqKey="Hoffmeister P" first="Paula" last="Hoffmeister">Paula Hoffmeister</name>
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<author>
<name sortKey="Neukirch, Lasse" sort="Neukirch, Lasse" uniqKey="Neukirch L" first="Lasse" last="Neukirch">Lasse Neukirch</name>
<affiliation>
<nlm:aff id="af2-ijms-21-01099">Clinical Cooperation Unit Applied Tumor Immunity, National Center for Tumor Diseases and German Cancer Research Center, Heidelberg 69120, Germany;
<email>lasse.neukirch@nct-heidelberg.de</email>
</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey="Gdynia, Georg" sort="Gdynia, Georg" uniqKey="Gdynia G" first="Georg" last="Gdynia">Georg Gdynia</name>
<affiliation>
<nlm:aff id="af3-ijms-21-01099">Institute of Pathology, University Hospital Heidelberg, Heidelberg 69120, Germany;
<email>Georg.Gdynia@med.uni-heidelberg.de</email>
(G.G.);
<email>Benjamin.Goeppert@med.uni-heidelberg.de</email>
(B.G.)</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey="Goeppert, Benjamin" sort="Goeppert, Benjamin" uniqKey="Goeppert B" first="Benjamin" last="Goeppert">Benjamin Goeppert</name>
<affiliation>
<nlm:aff id="af3-ijms-21-01099">Institute of Pathology, University Hospital Heidelberg, Heidelberg 69120, Germany;
<email>Georg.Gdynia@med.uni-heidelberg.de</email>
(G.G.);
<email>Benjamin.Goeppert@med.uni-heidelberg.de</email>
(B.G.)</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey="Schulze Bergkamen, Henning" sort="Schulze Bergkamen, Henning" uniqKey="Schulze Bergkamen H" first="Henning" last="Schulze-Bergkamen">Henning Schulze-Bergkamen</name>
<affiliation>
<nlm:aff id="af4-ijms-21-01099">Department of Internal Medicine II, Marien Hospital, Wesel46483, Germany;
<email>Henning.Schulze-Bergkamen@prohomine.de</email>
</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey="J Ger, Dirk" sort="J Ger, Dirk" uniqKey="J Ger D" first="Dirk" last="J Ger">Dirk J Ger</name>
<affiliation>
<nlm:aff id="af1-ijms-21-01099">National Center for Tumor Diseases, University Hospital Heidelberg, Heidelberg 69120, Germany;
<email>anna-lena.scherr@nct-heidelberg.de</email>
(A.-L.S.);
<email>adam@jassowicz.com</email>
(A.J.);
<email>patoannaterezia@gmail.com</email>
(A.P.);
<email>Christin.Elssner@gmx.de</email>
(C.E.);
<email>lars.ismail1986@googlemail.com</email>
(L.I.);
<email>nathalie.schmitt@nct-heidelberg.de</email>
(N.S.);
<email>paula.hoffmeister@googlemail.com</email>
(P.H.);
<email>dirk.jaeger@nct-heidelberg.de</email>
(D.J.)</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey="Kohler, Bruno Christian" sort="Kohler, Bruno Christian" uniqKey="Kohler B" first="Bruno Christian" last="Köhler">Bruno Christian Köhler</name>
<affiliation>
<nlm:aff id="af1-ijms-21-01099">National Center for Tumor Diseases, University Hospital Heidelberg, Heidelberg 69120, Germany;
<email>anna-lena.scherr@nct-heidelberg.de</email>
(A.-L.S.);
<email>adam@jassowicz.com</email>
(A.J.);
<email>patoannaterezia@gmail.com</email>
(A.P.);
<email>Christin.Elssner@gmx.de</email>
(C.E.);
<email>lars.ismail1986@googlemail.com</email>
(L.I.);
<email>nathalie.schmitt@nct-heidelberg.de</email>
(N.S.);
<email>paula.hoffmeister@googlemail.com</email>
(P.H.);
<email>dirk.jaeger@nct-heidelberg.de</email>
(D.J.)</nlm:aff>
</affiliation>
</author>
</analytic>
<series>
<title level="j">International Journal of Molecular Sciences</title>
<idno type="eISSN">1422-0067</idno>
<imprint>
<date when="2020">2020</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc>
<textClass></textClass>
</profileDesc>
</teiHeader>
<front>
<div type="abstract" xml:lang="en">
<p>Autophagy is a catabolic process that enables cells to degrade obsolete content and refuel energy depots. In colorectal cancer (CRC) autophagy has been shown to promote tumorigenesis through energy delivery in the condition of uncontrolled proliferation. With this study, we aimed at evaluating whether autophagy sustains CRC cell viability and if it impacts therapy resistance. Initially, a colorectal cancer tissue micro array, containing mucosa (
<italic>n</italic>
= 10), adenoma (
<italic>n</italic>
= 18) and adenocarcinoma (
<italic>n</italic>
= 49) spots, was stained for expression of essential autophagy proteins LC3b, Atg7, p62 and Beclin-1. Subsequently, central autophagy proteins were downregulated in CRC cells using siRNA technology. Viability assays, flow cytometry and immunoblotting were performed and three-dimensional cell culture was utilized to study autophagy in a tissue mimicking environment. In our study we found an upregulation of Atg7 in CRC. Furthermore, we identified Atg7 as crucial factor within the autophagy network for CRC cell viability. Its disruption induced cell death via triggering apoptosis and in combination with conventional chemotherapy it exerted synergistic effects in inducing CRC cell death. Cell death was strictly dependent on nuclear LC3b, since simultaneous knockdown of Atg7 and LC3b completely restored viability. This study unravels a novel cell death preventing function of Atg7 in interaction with LC3b, thereby unmasking a promising therapeutic target in CRC.</p>
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<name sortKey="Pat, Anna" sort="Pat, Anna" uniqKey="Pat A" first="Anna" last="Pat">Anna Pat</name>
<name sortKey="Scherr, Anna Lena" sort="Scherr, Anna Lena" uniqKey="Scherr A" first="Anna-Lena" last="Scherr">Anna-Lena Scherr</name>
<name sortKey="Schmitt, Nathalie" sort="Schmitt, Nathalie" uniqKey="Schmitt N" first="Nathalie" last="Schmitt">Nathalie Schmitt</name>
<name sortKey="Schulze Bergkamen, Henning" sort="Schulze Bergkamen, Henning" uniqKey="Schulze Bergkamen H" first="Henning" last="Schulze-Bergkamen">Henning Schulze-Bergkamen</name>
</noCountry>
</tree>
</affiliations>
</record>

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