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Identification of compound CA-5f as a novel late-stage autophagy inhibitor with potent anti-tumor effect against non-small cell lung cancer.

Identifieur interne : 000048 ( PubMed/Checkpoint ); précédent : 000047; suivant : 000049

Identification of compound CA-5f as a novel late-stage autophagy inhibitor with potent anti-tumor effect against non-small cell lung cancer.

Auteurs : Lu Zhang [République populaire de Chine] ; Pengfei Qiang [République populaire de Chine] ; Jingting Yu [République populaire de Chine] ; Yiming Miao [République populaire de Chine] ; Zhiqiang Chen [République populaire de Chine] ; Ju Qu [République populaire de Chine] ; Qianbing Zhao [République populaire de Chine] ; Zhuo Chen [République populaire de Chine] ; Yachao Liu [République populaire de Chine] ; Xin Yao [République populaire de Chine] ; Bin Liu [République populaire de Chine] ; Liuqing Cui [République populaire de Chine] ; Hongjuan Jing [République populaire de Chine] ; Gangchun Sun [République populaire de Chine]

Source :

RBID : pubmed:30145925

Abstract

Currently, particular focus is placed on the implication of autophagy in a variety of human diseases, including cancer. Discovery of small-molecule modulators of autophagy as well as their potential use as anti-cancer therapeutic agents would be of great significance. To this end, a series of curcumin analogs previously synthesized in our laboratory were screened. Among these compounds, (3E,5E)-3-(3,4-dimethoxybenzylidene)-5-[(1H-indol-3-yl)methylene]-1-methylpiperidin-4-one (CA-5f) was identified as a potent late-stage macroautophagy/autophagy inhibitor via inhibiting autophagosome-lysosome fusion. We found that CA-5f neither impaired the hydrolytic function nor the quantity of lysosomes. Use of an isobaric tag for relative and absolute quantitation (iTRAQ)-based proteomic screen in combination with bioinformatics analysis suggested that treatment of human umbilical vein endothelial cells (HUVECs) with CA-5f for 1 h suppressed the levels of cytoskeletal proteins and membrane traffic proteins. Subsequent studies showed that CA-5f exhibited strong cytotoxicity against A549 non-small cell lung cancer (NSCLC) cells, but low cytotoxicity to normal human umbilical vein endothelial cells (HUVECs), by increasing mitochondrial-derived reactive oxygen species (ROS) production. Moreover, CA-5f effectively suppressed the growth of A549 lung cancer xenograft as a single agent with an excellent tolerance in vivo. Results from western blot, immunofluorescence, and TdT-mediated dUTP nick end labeling (TUNEL) assays showed that CA-5f inhibited autophagic flux, induced apoptosis, and did not affect the level of CTSB (cathepsin B) and CTSD (cathepsin D) in vivo, which were consistent with the in vitro data. Collectively, these results demonstrated that CA-5f is a novel late-stage autophagy inhibitor with potential clinical application for NSCLC therapy. Abbreviations: 3-MA, 3-methyladenine; ANXA5, annexin A5; ATG, autophagy related; CA-5f, (3E,5E)-3-(3,4-dimethoxybenzylidene)-5-[(1H-indol-3-yl)methylene]-1-methylpiperidin-4-one; CQ, chloroquine; CTSB, cathepsin B; CTSD, cathepsin D; DMSO, dimethyl sulfoxide; DNM2, dynamin 2; EBSS, Earle's balanced salt solution; GFP, green fluorescent protein; HCQ, hydroxyl CQ; HEK293, human embryonic kidney 293; HUVEC, human umbilical vein endothelial cells; LAMP1, lysosomal associated membrane protein 1; LC-MS/MS, liquid chromatography coupled to tandem mass spectrometry; LDH, lactic acid dehydrogenase; LMO7, LIM domain 7; MAP1LC3B/LC3B, microtubule associated protein 1 light chain 3 beta; NAC, N-acetyl cysteine; MYO1E, myosin IE; NSCLC, non-small cell lung cancer; PARP1, poly(ADP-ribose) polymerase 1; PI, propidium iodide; RFP, red fluorescent protein; ROS, reactive oxygen species; SQSTM1, sequestosome 1; TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling.

DOI: 10.1080/15548627.2018.1511503
PubMed: 30145925


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<div type="abstract" xml:lang="en">Currently, particular focus is placed on the implication of autophagy in a variety of human diseases, including cancer. Discovery of small-molecule modulators of autophagy as well as their potential use as anti-cancer therapeutic agents would be of great significance. To this end, a series of curcumin analogs previously synthesized in our laboratory were screened. Among these compounds, (3E,5E)-3-(3,4-dimethoxybenzylidene)-5-[(1H-indol-3-yl)methylene]-1-methylpiperidin-4-one (CA-5f) was identified as a potent late-stage macroautophagy/autophagy inhibitor via inhibiting autophagosome-lysosome fusion. We found that CA-5f neither impaired the hydrolytic function nor the quantity of lysosomes. Use of an isobaric tag for relative and absolute quantitation (iTRAQ)-based proteomic screen in combination with bioinformatics analysis suggested that treatment of human umbilical vein endothelial cells (HUVECs) with CA-5f for 1 h suppressed the levels of cytoskeletal proteins and membrane traffic proteins. Subsequent studies showed that CA-5f exhibited strong cytotoxicity against A549 non-small cell lung cancer (NSCLC) cells, but low cytotoxicity to normal human umbilical vein endothelial cells (HUVECs), by increasing mitochondrial-derived reactive oxygen species (ROS) production. Moreover, CA-5f effectively suppressed the growth of A549 lung cancer xenograft as a single agent with an excellent tolerance in vivo. Results from western blot, immunofluorescence, and TdT-mediated dUTP nick end labeling (TUNEL) assays showed that CA-5f inhibited autophagic flux, induced apoptosis, and did not affect the level of CTSB (cathepsin B) and CTSD (cathepsin D) in vivo, which were consistent with the in vitro data. Collectively, these results demonstrated that CA-5f is a novel late-stage autophagy inhibitor with potential clinical application for NSCLC therapy. Abbreviations: 3-MA, 3-methyladenine; ANXA5, annexin A5; ATG, autophagy related; CA-5f, (3E,5E)-3-(3,4-dimethoxybenzylidene)-5-[(1H-indol-3-yl)methylene]-1-methylpiperidin-4-one; CQ, chloroquine; CTSB, cathepsin B; CTSD, cathepsin D; DMSO, dimethyl sulfoxide; DNM2, dynamin 2; EBSS, Earle's balanced salt solution; GFP, green fluorescent protein; HCQ, hydroxyl CQ; HEK293, human embryonic kidney 293; HUVEC, human umbilical vein endothelial cells; LAMP1, lysosomal associated membrane protein 1; LC-MS/MS, liquid chromatography coupled to tandem mass spectrometry; LDH, lactic acid dehydrogenase; LMO7, LIM domain 7; MAP1LC3B/LC3B, microtubule associated protein 1 light chain 3 beta; NAC, N-acetyl cysteine; MYO1E, myosin IE; NSCLC, non-small cell lung cancer; PARP1, poly(ADP-ribose) polymerase 1; PI, propidium iodide; RFP, red fluorescent protein; ROS, reactive oxygen species; SQSTM1, sequestosome 1; TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling.</div>
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<Abstract>
<AbstractText>Currently, particular focus is placed on the implication of autophagy in a variety of human diseases, including cancer. Discovery of small-molecule modulators of autophagy as well as their potential use as anti-cancer therapeutic agents would be of great significance. To this end, a series of curcumin analogs previously synthesized in our laboratory were screened. Among these compounds, (3E,5E)-3-(3,4-dimethoxybenzylidene)-5-[(1H-indol-3-yl)methylene]-1-methylpiperidin-4-one (CA-5f) was identified as a potent late-stage macroautophagy/autophagy inhibitor via inhibiting autophagosome-lysosome fusion. We found that CA-5f neither impaired the hydrolytic function nor the quantity of lysosomes. Use of an isobaric tag for relative and absolute quantitation (iTRAQ)-based proteomic screen in combination with bioinformatics analysis suggested that treatment of human umbilical vein endothelial cells (HUVECs) with CA-5f for 1 h suppressed the levels of cytoskeletal proteins and membrane traffic proteins. Subsequent studies showed that CA-5f exhibited strong cytotoxicity against A549 non-small cell lung cancer (NSCLC) cells, but low cytotoxicity to normal human umbilical vein endothelial cells (HUVECs), by increasing mitochondrial-derived reactive oxygen species (ROS) production. Moreover, CA-5f effectively suppressed the growth of A549 lung cancer xenograft as a single agent with an excellent tolerance in vivo. Results from western blot, immunofluorescence, and TdT-mediated dUTP nick end labeling (TUNEL) assays showed that CA-5f inhibited autophagic flux, induced apoptosis, and did not affect the level of CTSB (cathepsin B) and CTSD (cathepsin D) in vivo, which were consistent with the in vitro data. Collectively, these results demonstrated that CA-5f is a novel late-stage autophagy inhibitor with potential clinical application for NSCLC therapy. Abbreviations: 3-MA, 3-methyladenine; ANXA5, annexin A5; ATG, autophagy related; CA-5f, (3E,5E)-3-(3,4-dimethoxybenzylidene)-5-[(1H-indol-3-yl)methylene]-1-methylpiperidin-4-one; CQ, chloroquine; CTSB, cathepsin B; CTSD, cathepsin D; DMSO, dimethyl sulfoxide; DNM2, dynamin 2; EBSS, Earle's balanced salt solution; GFP, green fluorescent protein; HCQ, hydroxyl CQ; HEK293, human embryonic kidney 293; HUVEC, human umbilical vein endothelial cells; LAMP1, lysosomal associated membrane protein 1; LC-MS/MS, liquid chromatography coupled to tandem mass spectrometry; LDH, lactic acid dehydrogenase; LMO7, LIM domain 7; MAP1LC3B/LC3B, microtubule associated protein 1 light chain 3 beta; NAC, N-acetyl cysteine; MYO1E, myosin IE; NSCLC, non-small cell lung cancer; PARP1, poly(ADP-ribose) polymerase 1; PI, propidium iodide; RFP, red fluorescent protein; ROS, reactive oxygen species; SQSTM1, sequestosome 1; TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling.</AbstractText>
</Abstract>
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<LastName>Zhang</LastName>
<ForeName>Lu</ForeName>
<Initials>L</Initials>
<AffiliationInfo>
<Affiliation>a College of Bioengineering , Henan University of Technology , Zhengzhou , China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y">
<LastName>Qiang</LastName>
<ForeName>PengFei</ForeName>
<Initials>P</Initials>
<AffiliationInfo>
<Affiliation>a College of Bioengineering , Henan University of Technology , Zhengzhou , China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y">
<LastName>Yu</LastName>
<ForeName>JingTing</ForeName>
<Initials>J</Initials>
<Identifier Source="ORCID">0000-0002-2631-8434</Identifier>
<AffiliationInfo>
<Affiliation>a College of Bioengineering , Henan University of Technology , Zhengzhou , China.</Affiliation>
</AffiliationInfo>
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<LastName>Miao</LastName>
<ForeName>YiMing</ForeName>
<Initials>Y</Initials>
<AffiliationInfo>
<Affiliation>a College of Bioengineering , Henan University of Technology , Zhengzhou , China.</Affiliation>
</AffiliationInfo>
</Author>
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<LastName>Chen</LastName>
<ForeName>ZhiQiang</ForeName>
<Initials>Z</Initials>
<AffiliationInfo>
<Affiliation>a College of Bioengineering , Henan University of Technology , Zhengzhou , China.</Affiliation>
</AffiliationInfo>
</Author>
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<LastName>Qu</LastName>
<ForeName>Ju</ForeName>
<Initials>J</Initials>
<AffiliationInfo>
<Affiliation>a College of Bioengineering , Henan University of Technology , Zhengzhou , China.</Affiliation>
</AffiliationInfo>
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<LastName>Zhao</LastName>
<ForeName>QianBing</ForeName>
<Initials>Q</Initials>
<AffiliationInfo>
<Affiliation>a College of Bioengineering , Henan University of Technology , Zhengzhou , China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y">
<LastName>Chen</LastName>
<ForeName>Zhuo</ForeName>
<Initials>Z</Initials>
<AffiliationInfo>
<Affiliation>a College of Bioengineering , Henan University of Technology , Zhengzhou , China.</Affiliation>
</AffiliationInfo>
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<ForeName>Yachao</ForeName>
<Initials>Y</Initials>
<AffiliationInfo>
<Affiliation>a College of Bioengineering , Henan University of Technology , Zhengzhou , China.</Affiliation>
</AffiliationInfo>
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<LastName>Yao</LastName>
<ForeName>Xin</ForeName>
<Initials>X</Initials>
<AffiliationInfo>
<Affiliation>a College of Bioengineering , Henan University of Technology , Zhengzhou , China.</Affiliation>
</AffiliationInfo>
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<LastName>Liu</LastName>
<ForeName>Bin</ForeName>
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<AffiliationInfo>
<Affiliation>a College of Bioengineering , Henan University of Technology , Zhengzhou , China.</Affiliation>
</AffiliationInfo>
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<LastName>Cui</LastName>
<ForeName>LiuQing</ForeName>
<Initials>L</Initials>
<AffiliationInfo>
<Affiliation>a College of Bioengineering , Henan University of Technology , Zhengzhou , China.</Affiliation>
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<Author ValidYN="Y">
<LastName>Jing</LastName>
<ForeName>HongJuan</ForeName>
<Initials>H</Initials>
<AffiliationInfo>
<Affiliation>a College of Bioengineering , Henan University of Technology , Zhengzhou , China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y">
<LastName>Sun</LastName>
<ForeName>Gangchun</ForeName>
<Initials>G</Initials>
<AffiliationInfo>
<Affiliation>b College of Chemistry and Chemical Engineering , Henan University of Technology , Zhengzhou , China.</Affiliation>
</AffiliationInfo>
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<Year>2018</Year>
<Month>09</Month>
<Day>06</Day>
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<Country>United States</Country>
<MedlineTA>Autophagy</MedlineTA>
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<ISSNLinking>1554-8627</ISSNLinking>
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<Keyword MajorTopicYN="Y">Autophagy inhibitor</Keyword>
<Keyword MajorTopicYN="Y">CA-5f</Keyword>
<Keyword MajorTopicYN="Y">cell death</Keyword>
<Keyword MajorTopicYN="Y">curcumin analogs</Keyword>
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<Keyword MajorTopicYN="Y">non-small cell lung cancer</Keyword>
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   |clé=     pubmed:30145925
   |texte=   Identification of compound CA-5f as a novel late-stage autophagy inhibitor with potent anti-tumor effect against non-small cell lung cancer.
}}

Pour générer des pages wiki

HfdIndexSelect -h $EXPLOR_AREA/Data/PubMed/Checkpoint/RBID.i   -Sk "pubmed:30145925" \
       | HfdSelect -Kh $EXPLOR_AREA/Data/PubMed/Checkpoint/biblio.hfd   \
       | NlmPubMed2Wicri -a ChloroquineV1 

Wicri

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