Induction of ROS-Dependent Mitochondria-Mediated Intrinsic Apoptosis in MDA-MB-231 Cells by Glycoprotein from Codium decorticatum
Marine macroalgae consist of a range of bioactive molecules exhibiting different biological activities, and many of these properties are attributed to sulfated polysaccharides, fucoxanthin, phycobiliproteins, and halogenated compounds. In this study, a glycoprotein (GLP) with a molecular mass of ∼48...
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| Veröffentlicht in: | Journal of agricultural and food chemistry 2014-04, Vol.62 (15), p.3410-3421 |
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| creator | Thangam, Ramar Senthilkumar, Dharmaraj Suresh, Veeraperumal Sathuvan, Malairaj Sivasubramanian, Srinivasan Pazhanichamy, Kalailingam Gorlagunta, Praveen Kumar Kannan, Soundarapandian Gunasekaran, Palani Rengasamy, Ramasamy Sivaraman, Jayanthi |
| description | Marine macroalgae consist of a range of bioactive molecules exhibiting different biological activities, and many of these properties are attributed to sulfated polysaccharides, fucoxanthin, phycobiliproteins, and halogenated compounds. In this study, a glycoprotein (GLP) with a molecular mass of ∼48 kDa was extracted and purified from Codium decorticatum and investigated for its cytotoxic properties against human MDA-MB-231 breast cancer cells. The IC50 values of GLP against MDA-MB-231 and normal breast HBL-100 cells (control) were 75 ± 0.23 μg/mL (IC25), 55 ± 0.32 μg/mL (IC50), and 30 ± 0.43 μg/mL (IC75) and 90 ± 0.57 μg/mL (IC25), 80 ± 0.48 μg/mL (IC50), and 60 ± 0.26 μg/mL (IC75), respectively. Chromatin condensation and poly(ADP-ribose) polymerase (PARP) cleavage studies showed that the GLP inhibited cell viability by inducing apoptosis in MDA-MB-231 cells. Induction of mitochondria-mediated intrinsic apoptotic pathway by GLP was evidenced by the events of loss of mitochondrial membrane potential (ΔΨm), bax/bcl-2 dysregulation, cytochrome c release, and activation of caspases 3 and 9. Apoptosis-associated factors such as reactive oxygen species (ROS) formation and loss of ΔΨm were evaluated by DCFH-DA staining and flow cytometry, respectively. Cell cycle arrest of G2/M phase and expression of apoptosis associated proteins were determined using flow cytometry and Western blotting, respectively. |
| doi_str_mv | 10.1021/jf405329e |
| format | Article |
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In this study, a glycoprotein (GLP) with a molecular mass of ∼48 kDa was extracted and purified from Codium decorticatum and investigated for its cytotoxic properties against human MDA-MB-231 breast cancer cells. The IC50 values of GLP against MDA-MB-231 and normal breast HBL-100 cells (control) were 75 ± 0.23 μg/mL (IC25), 55 ± 0.32 μg/mL (IC50), and 30 ± 0.43 μg/mL (IC75) and 90 ± 0.57 μg/mL (IC25), 80 ± 0.48 μg/mL (IC50), and 60 ± 0.26 μg/mL (IC75), respectively. Chromatin condensation and poly(ADP-ribose) polymerase (PARP) cleavage studies showed that the GLP inhibited cell viability by inducing apoptosis in MDA-MB-231 cells. Induction of mitochondria-mediated intrinsic apoptotic pathway by GLP was evidenced by the events of loss of mitochondrial membrane potential (ΔΨm), bax/bcl-2 dysregulation, cytochrome c release, and activation of caspases 3 and 9. Apoptosis-associated factors such as reactive oxygen species (ROS) formation and loss of ΔΨm were evaluated by DCFH-DA staining and flow cytometry, respectively. Cell cycle arrest of G2/M phase and expression of apoptosis associated proteins were determined using flow cytometry and Western blotting, respectively.</description><identifier>ISSN: 0021-8561</identifier><identifier>EISSN: 1520-5118</identifier><identifier>DOI: 10.1021/jf405329e</identifier><identifier>PMID: 24694116</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>apoptosis ; Apoptosis - drug effects ; bcl-2-Associated X Protein - genetics ; bcl-2-Associated X Protein - metabolism ; breast neoplasms ; caspases ; cell cycle checkpoints ; Cell Cycle Checkpoints - drug effects ; Cell Line, Tumor ; cell viability ; Chlorophyta - chemistry ; chromatin ; Codium ; cytochrome c ; cytotoxicity ; flow cytometry ; glycoproteins ; Glycoproteins - isolation & purification ; Glycoproteins - pharmacology ; Humans ; inhibitory concentration 50 ; macroalgae ; membrane potential ; Mitochondria - drug effects ; Mitochondria - metabolism ; mitochondrial membrane ; molecular weight ; NAD ADP-ribosyltransferase ; neoplasm cells ; phycobiliprotein ; polysaccharides ; reactive oxygen species ; Reactive Oxygen Species - metabolism ; Signal Transduction - drug effects ; Western blotting</subject><ispartof>Journal of agricultural and food chemistry, 2014-04, Vol.62 (15), p.3410-3421</ispartof><rights>Copyright © 2014 American Chemical Society</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a405t-fff26b8e0212eb4fa30c7ca519d02149b2baf62989801ab5a015be3fcbb02b783</citedby><cites>FETCH-LOGICAL-a405t-fff26b8e0212eb4fa30c7ca519d02149b2baf62989801ab5a015be3fcbb02b783</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/jf405329e$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/jf405329e$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,780,784,2765,27076,27924,27925,56738,56788</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24694116$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Thangam, Ramar</creatorcontrib><creatorcontrib>Senthilkumar, Dharmaraj</creatorcontrib><creatorcontrib>Suresh, Veeraperumal</creatorcontrib><creatorcontrib>Sathuvan, Malairaj</creatorcontrib><creatorcontrib>Sivasubramanian, Srinivasan</creatorcontrib><creatorcontrib>Pazhanichamy, Kalailingam</creatorcontrib><creatorcontrib>Gorlagunta, Praveen Kumar</creatorcontrib><creatorcontrib>Kannan, Soundarapandian</creatorcontrib><creatorcontrib>Gunasekaran, Palani</creatorcontrib><creatorcontrib>Rengasamy, Ramasamy</creatorcontrib><creatorcontrib>Sivaraman, Jayanthi</creatorcontrib><title>Induction of ROS-Dependent Mitochondria-Mediated Intrinsic Apoptosis in MDA-MB-231 Cells by Glycoprotein from Codium decorticatum</title><title>Journal of agricultural and food chemistry</title><addtitle>J. Agric. Food Chem</addtitle><description>Marine macroalgae consist of a range of bioactive molecules exhibiting different biological activities, and many of these properties are attributed to sulfated polysaccharides, fucoxanthin, phycobiliproteins, and halogenated compounds. In this study, a glycoprotein (GLP) with a molecular mass of ∼48 kDa was extracted and purified from Codium decorticatum and investigated for its cytotoxic properties against human MDA-MB-231 breast cancer cells. The IC50 values of GLP against MDA-MB-231 and normal breast HBL-100 cells (control) were 75 ± 0.23 μg/mL (IC25), 55 ± 0.32 μg/mL (IC50), and 30 ± 0.43 μg/mL (IC75) and 90 ± 0.57 μg/mL (IC25), 80 ± 0.48 μg/mL (IC50), and 60 ± 0.26 μg/mL (IC75), respectively. Chromatin condensation and poly(ADP-ribose) polymerase (PARP) cleavage studies showed that the GLP inhibited cell viability by inducing apoptosis in MDA-MB-231 cells. Induction of mitochondria-mediated intrinsic apoptotic pathway by GLP was evidenced by the events of loss of mitochondrial membrane potential (ΔΨm), bax/bcl-2 dysregulation, cytochrome c release, and activation of caspases 3 and 9. Apoptosis-associated factors such as reactive oxygen species (ROS) formation and loss of ΔΨm were evaluated by DCFH-DA staining and flow cytometry, respectively. Cell cycle arrest of G2/M phase and expression of apoptosis associated proteins were determined using flow cytometry and Western blotting, respectively.</description><subject>apoptosis</subject><subject>Apoptosis - drug effects</subject><subject>bcl-2-Associated X Protein - genetics</subject><subject>bcl-2-Associated X Protein - metabolism</subject><subject>breast neoplasms</subject><subject>caspases</subject><subject>cell cycle checkpoints</subject><subject>Cell Cycle Checkpoints - drug effects</subject><subject>Cell Line, Tumor</subject><subject>cell viability</subject><subject>Chlorophyta - chemistry</subject><subject>chromatin</subject><subject>Codium</subject><subject>cytochrome c</subject><subject>cytotoxicity</subject><subject>flow cytometry</subject><subject>glycoproteins</subject><subject>Glycoproteins - isolation & purification</subject><subject>Glycoproteins - pharmacology</subject><subject>Humans</subject><subject>inhibitory concentration 50</subject><subject>macroalgae</subject><subject>membrane potential</subject><subject>Mitochondria - drug effects</subject><subject>Mitochondria - metabolism</subject><subject>mitochondrial membrane</subject><subject>molecular weight</subject><subject>NAD ADP-ribosyltransferase</subject><subject>neoplasm cells</subject><subject>phycobiliprotein</subject><subject>polysaccharides</subject><subject>reactive oxygen species</subject><subject>Reactive Oxygen Species - metabolism</subject><subject>Signal Transduction - drug effects</subject><subject>Western blotting</subject><issn>0021-8561</issn><issn>1520-5118</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNptkEtvFDEQhC0EIkvgwB8AX5DgYOj2jOdxXDYPVsoqEiHnke2xwasde7A9hz3yz3G0ISdOLXV_qq4qQt4ifEbg-GVvaxAV780zskLBgQnE7jlZQTmyTjR4Rl6ltAeATrTwkpzxuulrxGZF_mz9uOjsgqfB0u-3d-zCzMaPxme6cznoX8GP0Um2M6OT2Yx063N0PjlN13OYc0guUefp7mLNdl8Zr5BuzOGQqDrS68NRhzmGbApgY5joJoxumehodIjZaZmX6TV5YeUhmTeP85zcX13-2HxjN7fX2836hskSLjNrLW9UZ0okblRtZQW61VJgP5ZV3SuupG143_UdoFRCAgplKquVAq7arjonH0-6xdDvxaQ8TC7pYlV6E5Y0oMC2BmhFXdBPJ1THkFI0dpijm2Q8DgjDQ-PDU-OFffcou6jJjE_kv4oL8P4EWBkG-TO6NNzfccAGALGv4cHZhxMhdRr2YYm-9PCfV38BQq-Q8g</recordid><startdate>20140416</startdate><enddate>20140416</enddate><creator>Thangam, Ramar</creator><creator>Senthilkumar, Dharmaraj</creator><creator>Suresh, Veeraperumal</creator><creator>Sathuvan, Malairaj</creator><creator>Sivasubramanian, Srinivasan</creator><creator>Pazhanichamy, Kalailingam</creator><creator>Gorlagunta, Praveen Kumar</creator><creator>Kannan, Soundarapandian</creator><creator>Gunasekaran, Palani</creator><creator>Rengasamy, Ramasamy</creator><creator>Sivaraman, Jayanthi</creator><general>American Chemical Society</general><general>American Chemical Society, Books and Journals Division</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20140416</creationdate><title>Induction of ROS-Dependent Mitochondria-Mediated Intrinsic Apoptosis in MDA-MB-231 Cells by Glycoprotein from Codium decorticatum</title><author>Thangam, Ramar ; Senthilkumar, Dharmaraj ; Suresh, Veeraperumal ; Sathuvan, Malairaj ; Sivasubramanian, Srinivasan ; Pazhanichamy, Kalailingam ; Gorlagunta, Praveen Kumar ; Kannan, Soundarapandian ; Gunasekaran, Palani ; Rengasamy, Ramasamy ; Sivaraman, Jayanthi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a405t-fff26b8e0212eb4fa30c7ca519d02149b2baf62989801ab5a015be3fcbb02b783</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>apoptosis</topic><topic>Apoptosis - drug effects</topic><topic>bcl-2-Associated X Protein - genetics</topic><topic>bcl-2-Associated X Protein - metabolism</topic><topic>breast neoplasms</topic><topic>caspases</topic><topic>cell cycle checkpoints</topic><topic>Cell Cycle Checkpoints - drug effects</topic><topic>Cell Line, Tumor</topic><topic>cell viability</topic><topic>Chlorophyta - chemistry</topic><topic>chromatin</topic><topic>Codium</topic><topic>cytochrome c</topic><topic>cytotoxicity</topic><topic>flow cytometry</topic><topic>glycoproteins</topic><topic>Glycoproteins - isolation & purification</topic><topic>Glycoproteins - pharmacology</topic><topic>Humans</topic><topic>inhibitory concentration 50</topic><topic>macroalgae</topic><topic>membrane potential</topic><topic>Mitochondria - drug effects</topic><topic>Mitochondria - metabolism</topic><topic>mitochondrial membrane</topic><topic>molecular weight</topic><topic>NAD ADP-ribosyltransferase</topic><topic>neoplasm cells</topic><topic>phycobiliprotein</topic><topic>polysaccharides</topic><topic>reactive oxygen species</topic><topic>Reactive Oxygen Species - metabolism</topic><topic>Signal Transduction - drug effects</topic><topic>Western blotting</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Thangam, Ramar</creatorcontrib><creatorcontrib>Senthilkumar, Dharmaraj</creatorcontrib><creatorcontrib>Suresh, Veeraperumal</creatorcontrib><creatorcontrib>Sathuvan, Malairaj</creatorcontrib><creatorcontrib>Sivasubramanian, Srinivasan</creatorcontrib><creatorcontrib>Pazhanichamy, Kalailingam</creatorcontrib><creatorcontrib>Gorlagunta, Praveen Kumar</creatorcontrib><creatorcontrib>Kannan, Soundarapandian</creatorcontrib><creatorcontrib>Gunasekaran, Palani</creatorcontrib><creatorcontrib>Rengasamy, Ramasamy</creatorcontrib><creatorcontrib>Sivaraman, Jayanthi</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of agricultural and food chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Thangam, Ramar</au><au>Senthilkumar, Dharmaraj</au><au>Suresh, Veeraperumal</au><au>Sathuvan, Malairaj</au><au>Sivasubramanian, Srinivasan</au><au>Pazhanichamy, Kalailingam</au><au>Gorlagunta, Praveen Kumar</au><au>Kannan, Soundarapandian</au><au>Gunasekaran, Palani</au><au>Rengasamy, Ramasamy</au><au>Sivaraman, Jayanthi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Induction of ROS-Dependent Mitochondria-Mediated Intrinsic Apoptosis in MDA-MB-231 Cells by Glycoprotein from Codium decorticatum</atitle><jtitle>Journal of agricultural and food chemistry</jtitle><addtitle>J. Agric. Food Chem</addtitle><date>2014-04-16</date><risdate>2014</risdate><volume>62</volume><issue>15</issue><spage>3410</spage><epage>3421</epage><pages>3410-3421</pages><issn>0021-8561</issn><eissn>1520-5118</eissn><abstract>Marine macroalgae consist of a range of bioactive molecules exhibiting different biological activities, and many of these properties are attributed to sulfated polysaccharides, fucoxanthin, phycobiliproteins, and halogenated compounds. In this study, a glycoprotein (GLP) with a molecular mass of ∼48 kDa was extracted and purified from Codium decorticatum and investigated for its cytotoxic properties against human MDA-MB-231 breast cancer cells. The IC50 values of GLP against MDA-MB-231 and normal breast HBL-100 cells (control) were 75 ± 0.23 μg/mL (IC25), 55 ± 0.32 μg/mL (IC50), and 30 ± 0.43 μg/mL (IC75) and 90 ± 0.57 μg/mL (IC25), 80 ± 0.48 μg/mL (IC50), and 60 ± 0.26 μg/mL (IC75), respectively. Chromatin condensation and poly(ADP-ribose) polymerase (PARP) cleavage studies showed that the GLP inhibited cell viability by inducing apoptosis in MDA-MB-231 cells. Induction of mitochondria-mediated intrinsic apoptotic pathway by GLP was evidenced by the events of loss of mitochondrial membrane potential (ΔΨm), bax/bcl-2 dysregulation, cytochrome c release, and activation of caspases 3 and 9. Apoptosis-associated factors such as reactive oxygen species (ROS) formation and loss of ΔΨm were evaluated by DCFH-DA staining and flow cytometry, respectively. Cell cycle arrest of G2/M phase and expression of apoptosis associated proteins were determined using flow cytometry and Western blotting, respectively.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>24694116</pmid><doi>10.1021/jf405329e</doi><tpages>12</tpages></addata></record> |
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| subjects | apoptosis Apoptosis - drug effects bcl-2-Associated X Protein - genetics bcl-2-Associated X Protein - metabolism breast neoplasms caspases cell cycle checkpoints Cell Cycle Checkpoints - drug effects Cell Line, Tumor cell viability Chlorophyta - chemistry chromatin Codium cytochrome c cytotoxicity flow cytometry glycoproteins Glycoproteins - isolation & purification Glycoproteins - pharmacology Humans inhibitory concentration 50 macroalgae membrane potential Mitochondria - drug effects Mitochondria - metabolism mitochondrial membrane molecular weight NAD ADP-ribosyltransferase neoplasm cells phycobiliprotein polysaccharides reactive oxygen species Reactive Oxygen Species - metabolism Signal Transduction - drug effects Western blotting |
| title | Induction of ROS-Dependent Mitochondria-Mediated Intrinsic Apoptosis in MDA-MB-231 Cells by Glycoprotein from Codium decorticatum |
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