Hsp90 inhibitor gedunin causes apoptosis in A549 lung cancer cells by disrupting Hsp90:Beclin-1:Bcl-2 interaction and downregulating autophagy
Hsp90 is regarded as an important therapeutic target in cancer treatment. Client proteins of Hsp90 like Beclin-1, PI3K, and AKT, are associated with tumor development, poor prognosis, and resistance to cancer therapies. This study aims to analyze the role of Gedunin, an Hsp-90 inhibitor, in mediatio...
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| Veröffentlicht in: | Life sciences (1973) 2020-09, Vol.256, p.118000, Article 118000 |
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| creator | Hasan, Adria Haque, Ejazul Hameed, Rohil Maier, Paul N. Irfan, Safia Kamil, Mohd Nazir, Aamir Mir, Snober S. |
| description | Hsp90 is regarded as an important therapeutic target in cancer treatment. Client proteins of Hsp90 like Beclin-1, PI3K, and AKT, are associated with tumor development, poor prognosis, and resistance to cancer therapies. This study aims to analyze the role of Gedunin, an Hsp-90 inhibitor, in mediation of crosstalk between apoptosis and autophagy by targeting Beclin-1:Bcl-2 interaction, and ER stress.
A549 cells were treated with different concentrations of gedunin, and inhibitory rate was evaluated by MTT assay. Effect of gedunin on generation of reactive oxygen species, mitochondrial membrane potential, and chromatin condensation was studied by staining methods like DCFH-DA, MitoTracker, and DAPI. Expression of EGFR, PIK3CA, AKT, marker genes for apoptosis and autophagy were studied using semi-quantitative RT-PCR. Interaction study of Hsp90:Beclin-1:Bcl-2 was done by immunoprecipitation analysis. Protein expression of autophagy and apoptosis markers along with Grp78, Hsp70, and Hsp90 was analyzed by immunoblotting.
Gedunin exerts cytotoxic effects, causes increase in ROS generation, downregulates mitochondrial membrane potential and induces loss in DNA integrity. mRNA expression analysis revealed that gedunin sensitized A549 cells towards apoptosis by downregulating EGFR, PIK3CA, AKT, and autophagy. Gedunin also inhibited interaction between Hsp90:Beclin-1:Bcl-2, leading to downregulation of autophagy (Beclin-1, Atg5-12 complex, and LC3) and antiapoptotic protein Bcl-2, which may result in ER stress-induced apoptosis. Moreover, Hsp90 inhibition by gedunin did not cause upregulation of Hsp70 expression.
Gedunin induces apoptosis in lung cancer cells by disrupting Hsp90:Beclin-1:Bcl-2 interaction and autophagy downregulation, thus making gedunin a good drug lead for targeting lung cancer.
•Hsp90 is an essential and highly abundant protein overexpressed in lung cancer.•Hsp90 client proteins help in cancer progression.•Hsp90 inhibitors targeting client proteins can help develop anti-cancer therapy.•Hsp90 inhibition caused autophagy inhibition by breaking Hsp90:Beclin-1:Bcl-2.•Lung cancer cells were chemo-sensitized towards ER-mediated apoptosis. |
| doi_str_mv | 10.1016/j.lfs.2020.118000 |
| format | Article |
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A549 cells were treated with different concentrations of gedunin, and inhibitory rate was evaluated by MTT assay. Effect of gedunin on generation of reactive oxygen species, mitochondrial membrane potential, and chromatin condensation was studied by staining methods like DCFH-DA, MitoTracker, and DAPI. Expression of EGFR, PIK3CA, AKT, marker genes for apoptosis and autophagy were studied using semi-quantitative RT-PCR. Interaction study of Hsp90:Beclin-1:Bcl-2 was done by immunoprecipitation analysis. Protein expression of autophagy and apoptosis markers along with Grp78, Hsp70, and Hsp90 was analyzed by immunoblotting.
Gedunin exerts cytotoxic effects, causes increase in ROS generation, downregulates mitochondrial membrane potential and induces loss in DNA integrity. mRNA expression analysis revealed that gedunin sensitized A549 cells towards apoptosis by downregulating EGFR, PIK3CA, AKT, and autophagy. Gedunin also inhibited interaction between Hsp90:Beclin-1:Bcl-2, leading to downregulation of autophagy (Beclin-1, Atg5-12 complex, and LC3) and antiapoptotic protein Bcl-2, which may result in ER stress-induced apoptosis. Moreover, Hsp90 inhibition by gedunin did not cause upregulation of Hsp70 expression.
Gedunin induces apoptosis in lung cancer cells by disrupting Hsp90:Beclin-1:Bcl-2 interaction and autophagy downregulation, thus making gedunin a good drug lead for targeting lung cancer.
•Hsp90 is an essential and highly abundant protein overexpressed in lung cancer.•Hsp90 client proteins help in cancer progression.•Hsp90 inhibitors targeting client proteins can help develop anti-cancer therapy.•Hsp90 inhibition caused autophagy inhibition by breaking Hsp90:Beclin-1:Bcl-2.•Lung cancer cells were chemo-sensitized towards ER-mediated apoptosis.</description><identifier>ISSN: 0024-3205</identifier><identifier>EISSN: 1879-0631</identifier><identifier>DOI: 10.1016/j.lfs.2020.118000</identifier><language>eng</language><publisher>New York: Elsevier Inc</publisher><subject>1-Phosphatidylinositol 3-kinase ; AKT protein ; Apoptosis ; Autophagy ; Bcl-2 protein ; Biomarkers ; Chromatin ; Crosstalk ; Cytotoxicity ; Deoxyribonucleic acid ; Disruption ; DNA ; Epidermal growth factor receptors ; ER stress ; Gedunin ; Gene expression ; Hsp70 protein ; Hsp90 ; Hsp90 protein ; Immunoblotting ; Immunoprecipitation ; Inhibitors ; Lung cancer ; Membrane potential ; Membranes ; Mitochondria ; Phagocytosis ; Polymerase chain reaction ; Proteins ; Reactive oxygen species ; Therapeutic targets</subject><ispartof>Life sciences (1973), 2020-09, Vol.256, p.118000, Article 118000</ispartof><rights>2020 Elsevier Inc.</rights><rights>Copyright Elsevier BV Sep 1, 2020</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c424t-799cd703dcb566b3e03ca8c2d3887ede04765f18ec0917ac647a98a120a0d8393</citedby><cites>FETCH-LOGICAL-c424t-799cd703dcb566b3e03ca8c2d3887ede04765f18ec0917ac647a98a120a0d8393</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.lfs.2020.118000$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids></links><search><creatorcontrib>Hasan, Adria</creatorcontrib><creatorcontrib>Haque, Ejazul</creatorcontrib><creatorcontrib>Hameed, Rohil</creatorcontrib><creatorcontrib>Maier, Paul N.</creatorcontrib><creatorcontrib>Irfan, Safia</creatorcontrib><creatorcontrib>Kamil, Mohd</creatorcontrib><creatorcontrib>Nazir, Aamir</creatorcontrib><creatorcontrib>Mir, Snober S.</creatorcontrib><title>Hsp90 inhibitor gedunin causes apoptosis in A549 lung cancer cells by disrupting Hsp90:Beclin-1:Bcl-2 interaction and downregulating autophagy</title><title>Life sciences (1973)</title><description>Hsp90 is regarded as an important therapeutic target in cancer treatment. Client proteins of Hsp90 like Beclin-1, PI3K, and AKT, are associated with tumor development, poor prognosis, and resistance to cancer therapies. This study aims to analyze the role of Gedunin, an Hsp-90 inhibitor, in mediation of crosstalk between apoptosis and autophagy by targeting Beclin-1:Bcl-2 interaction, and ER stress.
A549 cells were treated with different concentrations of gedunin, and inhibitory rate was evaluated by MTT assay. Effect of gedunin on generation of reactive oxygen species, mitochondrial membrane potential, and chromatin condensation was studied by staining methods like DCFH-DA, MitoTracker, and DAPI. Expression of EGFR, PIK3CA, AKT, marker genes for apoptosis and autophagy were studied using semi-quantitative RT-PCR. Interaction study of Hsp90:Beclin-1:Bcl-2 was done by immunoprecipitation analysis. Protein expression of autophagy and apoptosis markers along with Grp78, Hsp70, and Hsp90 was analyzed by immunoblotting.
Gedunin exerts cytotoxic effects, causes increase in ROS generation, downregulates mitochondrial membrane potential and induces loss in DNA integrity. mRNA expression analysis revealed that gedunin sensitized A549 cells towards apoptosis by downregulating EGFR, PIK3CA, AKT, and autophagy. Gedunin also inhibited interaction between Hsp90:Beclin-1:Bcl-2, leading to downregulation of autophagy (Beclin-1, Atg5-12 complex, and LC3) and antiapoptotic protein Bcl-2, which may result in ER stress-induced apoptosis. Moreover, Hsp90 inhibition by gedunin did not cause upregulation of Hsp70 expression.
Gedunin induces apoptosis in lung cancer cells by disrupting Hsp90:Beclin-1:Bcl-2 interaction and autophagy downregulation, thus making gedunin a good drug lead for targeting lung cancer.
•Hsp90 is an essential and highly abundant protein overexpressed in lung cancer.•Hsp90 client proteins help in cancer progression.•Hsp90 inhibitors targeting client proteins can help develop anti-cancer therapy.•Hsp90 inhibition caused autophagy inhibition by breaking Hsp90:Beclin-1:Bcl-2.•Lung cancer cells were chemo-sensitized towards ER-mediated apoptosis.</description><subject>1-Phosphatidylinositol 3-kinase</subject><subject>AKT protein</subject><subject>Apoptosis</subject><subject>Autophagy</subject><subject>Bcl-2 protein</subject><subject>Biomarkers</subject><subject>Chromatin</subject><subject>Crosstalk</subject><subject>Cytotoxicity</subject><subject>Deoxyribonucleic acid</subject><subject>Disruption</subject><subject>DNA</subject><subject>Epidermal growth factor receptors</subject><subject>ER stress</subject><subject>Gedunin</subject><subject>Gene expression</subject><subject>Hsp70 protein</subject><subject>Hsp90</subject><subject>Hsp90 protein</subject><subject>Immunoblotting</subject><subject>Immunoprecipitation</subject><subject>Inhibitors</subject><subject>Lung cancer</subject><subject>Membrane potential</subject><subject>Membranes</subject><subject>Mitochondria</subject><subject>Phagocytosis</subject><subject>Polymerase chain reaction</subject><subject>Proteins</subject><subject>Reactive oxygen species</subject><subject>Therapeutic targets</subject><issn>0024-3205</issn><issn>1879-0631</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><recordid>eNp9kMFOHDEMhqOqSN3SPkBvkXqexUkmkwk9AWoBCYlLOUfZxLtkNU2mSabVvgTPTGA5c7Is_59tfYR8Y7BmwIaz_XraljUH3no2AsAHsmKj0h0Mgn0kKwDed4KD_EQ-l7JvASmVWJGnmzJroCE-hk2oKdMd-iWGSJ1dChZq5zTXVEJpEXohe02nJe7aNDrM1OE0Fbo5UB9KXuYa2uh14fkluinEjp1fuqnjDa6YrashRWqjpz79jxl3y2RfGbvUND_a3eELOdnaqeDXt3pKHn79_H11093dX99eXdx1rud97ZTWzisQ3m3kMGwEgnB2dNyLcVToEXo1yC0b0YFmyrqhV1aPlnGw4EehxSn5ftw75_R3wVLNPi05tpOGSxiUlJoPLcWOKZdTKRm3Zs7hj80Hw8C8aDd707SbF-3mqL0xP44Mtvf_BcymuIDNlg8ZXTU-hXfoZ1Twiwc</recordid><startdate>20200901</startdate><enddate>20200901</enddate><creator>Hasan, Adria</creator><creator>Haque, Ejazul</creator><creator>Hameed, Rohil</creator><creator>Maier, Paul N.</creator><creator>Irfan, Safia</creator><creator>Kamil, Mohd</creator><creator>Nazir, Aamir</creator><creator>Mir, Snober S.</creator><general>Elsevier Inc</general><general>Elsevier BV</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7U7</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>20200901</creationdate><title>Hsp90 inhibitor gedunin causes apoptosis in A549 lung cancer cells by disrupting Hsp90:Beclin-1:Bcl-2 interaction and downregulating autophagy</title><author>Hasan, Adria ; Haque, Ejazul ; Hameed, Rohil ; Maier, Paul N. ; Irfan, Safia ; Kamil, Mohd ; Nazir, Aamir ; Mir, Snober S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c424t-799cd703dcb566b3e03ca8c2d3887ede04765f18ec0917ac647a98a120a0d8393</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>1-Phosphatidylinositol 3-kinase</topic><topic>AKT protein</topic><topic>Apoptosis</topic><topic>Autophagy</topic><topic>Bcl-2 protein</topic><topic>Biomarkers</topic><topic>Chromatin</topic><topic>Crosstalk</topic><topic>Cytotoxicity</topic><topic>Deoxyribonucleic acid</topic><topic>Disruption</topic><topic>DNA</topic><topic>Epidermal growth factor receptors</topic><topic>ER stress</topic><topic>Gedunin</topic><topic>Gene expression</topic><topic>Hsp70 protein</topic><topic>Hsp90</topic><topic>Hsp90 protein</topic><topic>Immunoblotting</topic><topic>Immunoprecipitation</topic><topic>Inhibitors</topic><topic>Lung cancer</topic><topic>Membrane potential</topic><topic>Membranes</topic><topic>Mitochondria</topic><topic>Phagocytosis</topic><topic>Polymerase chain reaction</topic><topic>Proteins</topic><topic>Reactive oxygen species</topic><topic>Therapeutic targets</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hasan, Adria</creatorcontrib><creatorcontrib>Haque, Ejazul</creatorcontrib><creatorcontrib>Hameed, Rohil</creatorcontrib><creatorcontrib>Maier, Paul N.</creatorcontrib><creatorcontrib>Irfan, Safia</creatorcontrib><creatorcontrib>Kamil, Mohd</creatorcontrib><creatorcontrib>Nazir, Aamir</creatorcontrib><creatorcontrib>Mir, Snober S.</creatorcontrib><collection>CrossRef</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Life sciences (1973)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hasan, Adria</au><au>Haque, Ejazul</au><au>Hameed, Rohil</au><au>Maier, Paul N.</au><au>Irfan, Safia</au><au>Kamil, Mohd</au><au>Nazir, Aamir</au><au>Mir, Snober S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Hsp90 inhibitor gedunin causes apoptosis in A549 lung cancer cells by disrupting Hsp90:Beclin-1:Bcl-2 interaction and downregulating autophagy</atitle><jtitle>Life sciences (1973)</jtitle><date>2020-09-01</date><risdate>2020</risdate><volume>256</volume><spage>118000</spage><pages>118000-</pages><artnum>118000</artnum><issn>0024-3205</issn><eissn>1879-0631</eissn><abstract>Hsp90 is regarded as an important therapeutic target in cancer treatment. Client proteins of Hsp90 like Beclin-1, PI3K, and AKT, are associated with tumor development, poor prognosis, and resistance to cancer therapies. This study aims to analyze the role of Gedunin, an Hsp-90 inhibitor, in mediation of crosstalk between apoptosis and autophagy by targeting Beclin-1:Bcl-2 interaction, and ER stress.
A549 cells were treated with different concentrations of gedunin, and inhibitory rate was evaluated by MTT assay. Effect of gedunin on generation of reactive oxygen species, mitochondrial membrane potential, and chromatin condensation was studied by staining methods like DCFH-DA, MitoTracker, and DAPI. Expression of EGFR, PIK3CA, AKT, marker genes for apoptosis and autophagy were studied using semi-quantitative RT-PCR. Interaction study of Hsp90:Beclin-1:Bcl-2 was done by immunoprecipitation analysis. Protein expression of autophagy and apoptosis markers along with Grp78, Hsp70, and Hsp90 was analyzed by immunoblotting.
Gedunin exerts cytotoxic effects, causes increase in ROS generation, downregulates mitochondrial membrane potential and induces loss in DNA integrity. mRNA expression analysis revealed that gedunin sensitized A549 cells towards apoptosis by downregulating EGFR, PIK3CA, AKT, and autophagy. Gedunin also inhibited interaction between Hsp90:Beclin-1:Bcl-2, leading to downregulation of autophagy (Beclin-1, Atg5-12 complex, and LC3) and antiapoptotic protein Bcl-2, which may result in ER stress-induced apoptosis. Moreover, Hsp90 inhibition by gedunin did not cause upregulation of Hsp70 expression.
Gedunin induces apoptosis in lung cancer cells by disrupting Hsp90:Beclin-1:Bcl-2 interaction and autophagy downregulation, thus making gedunin a good drug lead for targeting lung cancer.
•Hsp90 is an essential and highly abundant protein overexpressed in lung cancer.•Hsp90 client proteins help in cancer progression.•Hsp90 inhibitors targeting client proteins can help develop anti-cancer therapy.•Hsp90 inhibition caused autophagy inhibition by breaking Hsp90:Beclin-1:Bcl-2.•Lung cancer cells were chemo-sensitized towards ER-mediated apoptosis.</abstract><cop>New York</cop><pub>Elsevier Inc</pub><doi>10.1016/j.lfs.2020.118000</doi></addata></record> |
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| subjects | 1-Phosphatidylinositol 3-kinase AKT protein Apoptosis Autophagy Bcl-2 protein Biomarkers Chromatin Crosstalk Cytotoxicity Deoxyribonucleic acid Disruption DNA Epidermal growth factor receptors ER stress Gedunin Gene expression Hsp70 protein Hsp90 Hsp90 protein Immunoblotting Immunoprecipitation Inhibitors Lung cancer Membrane potential Membranes Mitochondria Phagocytosis Polymerase chain reaction Proteins Reactive oxygen species Therapeutic targets |
| title | Hsp90 inhibitor gedunin causes apoptosis in A549 lung cancer cells by disrupting Hsp90:Beclin-1:Bcl-2 interaction and downregulating autophagy |
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