SRJ23, a new semisynthetic andrographolide derivative: in vitro growth inhibition and mechanisms of cell cycle arrest and apoptosis in prostate cancer cells
Purpose 3,19-(3-Chloro-4-fluorobenzylidene)andrographolide (SRJ23), a new semisynthetic derivative of andrographolide (AGP), exhibited selectivity against prostate cancer cells in the US National Cancer Institute (NCI) in vitro anti-cancer screen. Herein, we report the in vitro growth inhibition and...
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| Veröffentlicht in: | Cell biology and toxicology 2014-10, Vol.30 (5), p.269-288 |
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| creator | Wong, Hui Chyn Wong, Charng Choon Sagineedu, Sreenivasa Rao Loke, Seng Cheong Lajis, Nordin Haji Stanslas, Johnson |
| description | Purpose
3,19-(3-Chloro-4-fluorobenzylidene)andrographolide (SRJ23), a new semisynthetic derivative of andrographolide (AGP), exhibited selectivity against prostate cancer cells in the US National Cancer Institute (NCI) in vitro anti-cancer screen. Herein, we report the in vitro growth inhibition and mechanisms of cell cycle arrest and apoptosis induced by SRJ23.
Methods
3-(4,5-Dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used in assessing in vitro growth inhibition of compounds against prostate cancer (PC-3, DU-145 and LNCaP) and mouse macrophage (RAW 264.7) cell lines. Flow cytometry was utilised to analyse cell cycle distribution, whereas fluorescence microscopy was performed to determine morphological cell death. DNA fragmentation and annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry were done to confirm apoptosis induced by SRJ23. Quantitation of cell cycle and apoptotic regulatory proteins were determined by immunoblotting.
Results
AGP and SRJ23 selectively inhibited the growth of prostate cancer cells compared with RAW 264.7 cells at low micromolar concentrations; however, SRJ23 was more potent. Mechanistically, SRJ23-treated PC-3 cells displayed down-regulation of cyclin-dependent kinase (CDK) 1 without affecting levels of CDK4 and cyclin D1. However, SRJ23 induced down-regulation of CDK4 and cyclin D1 but without affecting CDK1 in DU145 and LNCaP cell lines. DNA histogram analysis revealed that the SRJ23 induced G2/M in PC-3 cells but G1 arrest in DU-145 and LNCaP cells. Morphologically, both compounds induced predominantly apoptosis, which was further confirmed by DNA fragmentation and annexin V-FITC staining. The DNA fragmentation was inhibited in the presence of caspase 8 inhibitor (Z-IETD-FMK). Apoptosis was associated with an increase in caspase 8 expression and activation. This thought to have induced cleavage of Bid into t-Bid. Additionally, increased expression and activation of caspase 9 and Bax proteins were apparent, with a concomitant down-regulation of Bcl-2 protein. Similar apoptosis cascade of events was observed in SRJ23-treated DU145 and LNCaP cell lines.
Conclusion
SRJ23 inhibited the growth of prostate cancer cells by inducing G
2
/M and G1 arrest via down-regulation of CDK1, and CDK4 and cyclin, respectively, and initiated caspase-8-mediated mitochondrial apoptosis. Taken together, these data support the potential of this compound as a new anti-prostate cancer agent. |
| doi_str_mv | 10.1007/s10565-014-9282-5 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1618155351</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>3437560331</sourcerecordid><originalsourceid>FETCH-LOGICAL-c475t-2cc05d2ec5f84ef3264d89047e0e980a05055b3e3f01249f87b3a68066f98f8d3</originalsourceid><addsrcrecordid>eNp1kc2KFDEUhQtRnHb0AdxIwI0LS2-SSirlbhj8ZUDwZx3SqVtdGaqSMkn30O_iw5qaHkUEV-GS75x7klNVTym8ogDt60RBSFEDbeqOKVaLe9WGipbXUjF2v9pA27CaQUfPqkcpXQOApK14WJ0xAS0o3myqn1-_fGL8JTHE4w1JOLt09HnE7Cwxvo9hF80yhsn1SHqM7mCyO-Ab4jw5uBwD2cVwk8cyj27rsgt-lZEZ7Wi8S3MiYSAWp4nYo52QmBgx5VvGLGHJIbm0mi0xpGwyEmu8xXgrSY-rB4OZEj65O8-r7-_efrv8UF99fv_x8uKqtk0rcs2sBdEztGJQDQ6cyaZXHTQtAnYKDAgQYsuRD0BZ0w2q3XIjFUg5dGpQPT-vXpx8S4of-5JPl29YExiPYZ80lVRRIbigBX3-D3od9tGXdJoKyVuhJINC0RNly7NSxEEv0c0mHjUFvVanT9XpUp1eq9OiaJ7dOe-3M_Z_FL-7KgA7Aalc-R3Gv1b_1_UXEfyleg</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1563758620</pqid></control><display><type>article</type><title>SRJ23, a new semisynthetic andrographolide derivative: in vitro growth inhibition and mechanisms of cell cycle arrest and apoptosis in prostate cancer cells</title><source>MEDLINE</source><source>SpringerNature Complete Journals</source><creator>Wong, Hui Chyn ; Wong, Charng Choon ; Sagineedu, Sreenivasa Rao ; Loke, Seng Cheong ; Lajis, Nordin Haji ; Stanslas, Johnson</creator><creatorcontrib>Wong, Hui Chyn ; Wong, Charng Choon ; Sagineedu, Sreenivasa Rao ; Loke, Seng Cheong ; Lajis, Nordin Haji ; Stanslas, Johnson</creatorcontrib><description>Purpose
3,19-(3-Chloro-4-fluorobenzylidene)andrographolide (SRJ23), a new semisynthetic derivative of andrographolide (AGP), exhibited selectivity against prostate cancer cells in the US National Cancer Institute (NCI) in vitro anti-cancer screen. Herein, we report the in vitro growth inhibition and mechanisms of cell cycle arrest and apoptosis induced by SRJ23.
Methods
3-(4,5-Dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used in assessing in vitro growth inhibition of compounds against prostate cancer (PC-3, DU-145 and LNCaP) and mouse macrophage (RAW 264.7) cell lines. Flow cytometry was utilised to analyse cell cycle distribution, whereas fluorescence microscopy was performed to determine morphological cell death. DNA fragmentation and annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry were done to confirm apoptosis induced by SRJ23. Quantitation of cell cycle and apoptotic regulatory proteins were determined by immunoblotting.
Results
AGP and SRJ23 selectively inhibited the growth of prostate cancer cells compared with RAW 264.7 cells at low micromolar concentrations; however, SRJ23 was more potent. Mechanistically, SRJ23-treated PC-3 cells displayed down-regulation of cyclin-dependent kinase (CDK) 1 without affecting levels of CDK4 and cyclin D1. However, SRJ23 induced down-regulation of CDK4 and cyclin D1 but without affecting CDK1 in DU145 and LNCaP cell lines. DNA histogram analysis revealed that the SRJ23 induced G2/M in PC-3 cells but G1 arrest in DU-145 and LNCaP cells. Morphologically, both compounds induced predominantly apoptosis, which was further confirmed by DNA fragmentation and annexin V-FITC staining. The DNA fragmentation was inhibited in the presence of caspase 8 inhibitor (Z-IETD-FMK). Apoptosis was associated with an increase in caspase 8 expression and activation. This thought to have induced cleavage of Bid into t-Bid. Additionally, increased expression and activation of caspase 9 and Bax proteins were apparent, with a concomitant down-regulation of Bcl-2 protein. Similar apoptosis cascade of events was observed in SRJ23-treated DU145 and LNCaP cell lines.
Conclusion
SRJ23 inhibited the growth of prostate cancer cells by inducing G
2
/M and G1 arrest via down-regulation of CDK1, and CDK4 and cyclin, respectively, and initiated caspase-8-mediated mitochondrial apoptosis. Taken together, these data support the potential of this compound as a new anti-prostate cancer agent.</description><identifier>ISSN: 0742-2091</identifier><identifier>EISSN: 1573-6822</identifier><identifier>DOI: 10.1007/s10565-014-9282-5</identifier><identifier>PMID: 25070834</identifier><language>eng</language><publisher>Dordrecht: Springer Netherlands</publisher><subject>Animals ; Apoptosis ; Apoptosis - drug effects ; bcl-2-Associated X Protein - metabolism ; Biochemistry ; Biomedical and Life Sciences ; Caspase 8 - metabolism ; Caspase 9 - metabolism ; Cell Biology ; Cell cycle ; Cell Cycle Checkpoints - drug effects ; Cell Line, Tumor ; Cell Proliferation - drug effects ; Cyclin D1 - metabolism ; Cyclin-Dependent Kinases - metabolism ; Cytotoxicity ; Deoxyribonucleic acid ; Diterpenes - pharmacology ; DNA ; DNA Fragmentation - drug effects ; Down-Regulation - drug effects ; Fluorescence microscopy ; Guinea Pigs ; Humans ; Inhibitor drugs ; Iodides ; Life Sciences ; Male ; Original Research ; Pharmacology/Toxicology ; Prostate cancer ; Prostatic Neoplasms - drug therapy ; Prostatic Neoplasms - metabolism ; Proto-Oncogene Proteins c-bcl-2 - metabolism</subject><ispartof>Cell biology and toxicology, 2014-10, Vol.30 (5), p.269-288</ispartof><rights>Springer Science+Business Media Dordrecht 2014</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c475t-2cc05d2ec5f84ef3264d89047e0e980a05055b3e3f01249f87b3a68066f98f8d3</citedby><cites>FETCH-LOGICAL-c475t-2cc05d2ec5f84ef3264d89047e0e980a05055b3e3f01249f87b3a68066f98f8d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s10565-014-9282-5$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s10565-014-9282-5$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25070834$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wong, Hui Chyn</creatorcontrib><creatorcontrib>Wong, Charng Choon</creatorcontrib><creatorcontrib>Sagineedu, Sreenivasa Rao</creatorcontrib><creatorcontrib>Loke, Seng Cheong</creatorcontrib><creatorcontrib>Lajis, Nordin Haji</creatorcontrib><creatorcontrib>Stanslas, Johnson</creatorcontrib><title>SRJ23, a new semisynthetic andrographolide derivative: in vitro growth inhibition and mechanisms of cell cycle arrest and apoptosis in prostate cancer cells</title><title>Cell biology and toxicology</title><addtitle>Cell Biol Toxicol</addtitle><addtitle>Cell Biol Toxicol</addtitle><description>Purpose
3,19-(3-Chloro-4-fluorobenzylidene)andrographolide (SRJ23), a new semisynthetic derivative of andrographolide (AGP), exhibited selectivity against prostate cancer cells in the US National Cancer Institute (NCI) in vitro anti-cancer screen. Herein, we report the in vitro growth inhibition and mechanisms of cell cycle arrest and apoptosis induced by SRJ23.
Methods
3-(4,5-Dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used in assessing in vitro growth inhibition of compounds against prostate cancer (PC-3, DU-145 and LNCaP) and mouse macrophage (RAW 264.7) cell lines. Flow cytometry was utilised to analyse cell cycle distribution, whereas fluorescence microscopy was performed to determine morphological cell death. DNA fragmentation and annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry were done to confirm apoptosis induced by SRJ23. Quantitation of cell cycle and apoptotic regulatory proteins were determined by immunoblotting.
Results
AGP and SRJ23 selectively inhibited the growth of prostate cancer cells compared with RAW 264.7 cells at low micromolar concentrations; however, SRJ23 was more potent. Mechanistically, SRJ23-treated PC-3 cells displayed down-regulation of cyclin-dependent kinase (CDK) 1 without affecting levels of CDK4 and cyclin D1. However, SRJ23 induced down-regulation of CDK4 and cyclin D1 but without affecting CDK1 in DU145 and LNCaP cell lines. DNA histogram analysis revealed that the SRJ23 induced G2/M in PC-3 cells but G1 arrest in DU-145 and LNCaP cells. Morphologically, both compounds induced predominantly apoptosis, which was further confirmed by DNA fragmentation and annexin V-FITC staining. The DNA fragmentation was inhibited in the presence of caspase 8 inhibitor (Z-IETD-FMK). Apoptosis was associated with an increase in caspase 8 expression and activation. This thought to have induced cleavage of Bid into t-Bid. Additionally, increased expression and activation of caspase 9 and Bax proteins were apparent, with a concomitant down-regulation of Bcl-2 protein. Similar apoptosis cascade of events was observed in SRJ23-treated DU145 and LNCaP cell lines.
Conclusion
SRJ23 inhibited the growth of prostate cancer cells by inducing G
2
/M and G1 arrest via down-regulation of CDK1, and CDK4 and cyclin, respectively, and initiated caspase-8-mediated mitochondrial apoptosis. Taken together, these data support the potential of this compound as a new anti-prostate cancer agent.</description><subject>Animals</subject><subject>Apoptosis</subject><subject>Apoptosis - drug effects</subject><subject>bcl-2-Associated X Protein - metabolism</subject><subject>Biochemistry</subject><subject>Biomedical and Life Sciences</subject><subject>Caspase 8 - metabolism</subject><subject>Caspase 9 - metabolism</subject><subject>Cell Biology</subject><subject>Cell cycle</subject><subject>Cell Cycle Checkpoints - drug effects</subject><subject>Cell Line, Tumor</subject><subject>Cell Proliferation - drug effects</subject><subject>Cyclin D1 - metabolism</subject><subject>Cyclin-Dependent Kinases - metabolism</subject><subject>Cytotoxicity</subject><subject>Deoxyribonucleic acid</subject><subject>Diterpenes - pharmacology</subject><subject>DNA</subject><subject>DNA Fragmentation - drug effects</subject><subject>Down-Regulation - drug effects</subject><subject>Fluorescence microscopy</subject><subject>Guinea Pigs</subject><subject>Humans</subject><subject>Inhibitor drugs</subject><subject>Iodides</subject><subject>Life Sciences</subject><subject>Male</subject><subject>Original Research</subject><subject>Pharmacology/Toxicology</subject><subject>Prostate cancer</subject><subject>Prostatic Neoplasms - drug therapy</subject><subject>Prostatic Neoplasms - metabolism</subject><subject>Proto-Oncogene Proteins c-bcl-2 - metabolism</subject><issn>0742-2091</issn><issn>1573-6822</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp1kc2KFDEUhQtRnHb0AdxIwI0LS2-SSirlbhj8ZUDwZx3SqVtdGaqSMkn30O_iw5qaHkUEV-GS75x7klNVTym8ogDt60RBSFEDbeqOKVaLe9WGipbXUjF2v9pA27CaQUfPqkcpXQOApK14WJ0xAS0o3myqn1-_fGL8JTHE4w1JOLt09HnE7Cwxvo9hF80yhsn1SHqM7mCyO-Ab4jw5uBwD2cVwk8cyj27rsgt-lZEZ7Wi8S3MiYSAWp4nYo52QmBgx5VvGLGHJIbm0mi0xpGwyEmu8xXgrSY-rB4OZEj65O8-r7-_efrv8UF99fv_x8uKqtk0rcs2sBdEztGJQDQ6cyaZXHTQtAnYKDAgQYsuRD0BZ0w2q3XIjFUg5dGpQPT-vXpx8S4of-5JPl29YExiPYZ80lVRRIbigBX3-D3od9tGXdJoKyVuhJINC0RNly7NSxEEv0c0mHjUFvVanT9XpUp1eq9OiaJ7dOe-3M_Z_FL-7KgA7Aalc-R3Gv1b_1_UXEfyleg</recordid><startdate>20141001</startdate><enddate>20141001</enddate><creator>Wong, Hui Chyn</creator><creator>Wong, Charng Choon</creator><creator>Sagineedu, Sreenivasa Rao</creator><creator>Loke, Seng Cheong</creator><creator>Lajis, Nordin Haji</creator><creator>Stanslas, Johnson</creator><general>Springer Netherlands</general><general>Springer Nature B.V</general><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>3V.</scope><scope>7RV</scope><scope>7TK</scope><scope>7TM</scope><scope>7U7</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB0</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>NAPCQ</scope><scope>P64</scope><scope>PATMY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PYCSY</scope><scope>Q9U</scope><scope>RC3</scope></search><sort><creationdate>20141001</creationdate><title>SRJ23, a new semisynthetic andrographolide derivative: in vitro growth inhibition and mechanisms of cell cycle arrest and apoptosis in prostate cancer cells</title><author>Wong, Hui Chyn ; Wong, Charng Choon ; Sagineedu, Sreenivasa Rao ; Loke, Seng Cheong ; Lajis, Nordin Haji ; Stanslas, Johnson</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c475t-2cc05d2ec5f84ef3264d89047e0e980a05055b3e3f01249f87b3a68066f98f8d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Animals</topic><topic>Apoptosis</topic><topic>Apoptosis - drug effects</topic><topic>bcl-2-Associated X Protein - metabolism</topic><topic>Biochemistry</topic><topic>Biomedical and Life Sciences</topic><topic>Caspase 8 - metabolism</topic><topic>Caspase 9 - metabolism</topic><topic>Cell Biology</topic><topic>Cell cycle</topic><topic>Cell Cycle Checkpoints - drug effects</topic><topic>Cell Line, Tumor</topic><topic>Cell Proliferation - drug effects</topic><topic>Cyclin D1 - metabolism</topic><topic>Cyclin-Dependent Kinases - metabolism</topic><topic>Cytotoxicity</topic><topic>Deoxyribonucleic acid</topic><topic>Diterpenes - pharmacology</topic><topic>DNA</topic><topic>DNA Fragmentation - drug effects</topic><topic>Down-Regulation - drug effects</topic><topic>Fluorescence microscopy</topic><topic>Guinea Pigs</topic><topic>Humans</topic><topic>Inhibitor drugs</topic><topic>Iodides</topic><topic>Life Sciences</topic><topic>Male</topic><topic>Original Research</topic><topic>Pharmacology/Toxicology</topic><topic>Prostate cancer</topic><topic>Prostatic Neoplasms - drug therapy</topic><topic>Prostatic Neoplasms - metabolism</topic><topic>Proto-Oncogene Proteins c-bcl-2 - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wong, Hui Chyn</creatorcontrib><creatorcontrib>Wong, Charng Choon</creatorcontrib><creatorcontrib>Sagineedu, Sreenivasa Rao</creatorcontrib><creatorcontrib>Loke, Seng Cheong</creatorcontrib><creatorcontrib>Lajis, Nordin Haji</creatorcontrib><creatorcontrib>Stanslas, Johnson</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Nursing & Allied Health Database</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Biological Science Database</collection><collection>Nursing & Allied Health Premium</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Environmental Science Collection</collection><collection>ProQuest Central Basic</collection><collection>Genetics Abstracts</collection><jtitle>Cell biology and toxicology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wong, Hui Chyn</au><au>Wong, Charng Choon</au><au>Sagineedu, Sreenivasa Rao</au><au>Loke, Seng Cheong</au><au>Lajis, Nordin Haji</au><au>Stanslas, Johnson</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>SRJ23, a new semisynthetic andrographolide derivative: in vitro growth inhibition and mechanisms of cell cycle arrest and apoptosis in prostate cancer cells</atitle><jtitle>Cell biology and toxicology</jtitle><stitle>Cell Biol Toxicol</stitle><addtitle>Cell Biol Toxicol</addtitle><date>2014-10-01</date><risdate>2014</risdate><volume>30</volume><issue>5</issue><spage>269</spage><epage>288</epage><pages>269-288</pages><issn>0742-2091</issn><eissn>1573-6822</eissn><abstract>Purpose
3,19-(3-Chloro-4-fluorobenzylidene)andrographolide (SRJ23), a new semisynthetic derivative of andrographolide (AGP), exhibited selectivity against prostate cancer cells in the US National Cancer Institute (NCI) in vitro anti-cancer screen. Herein, we report the in vitro growth inhibition and mechanisms of cell cycle arrest and apoptosis induced by SRJ23.
Methods
3-(4,5-Dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used in assessing in vitro growth inhibition of compounds against prostate cancer (PC-3, DU-145 and LNCaP) and mouse macrophage (RAW 264.7) cell lines. Flow cytometry was utilised to analyse cell cycle distribution, whereas fluorescence microscopy was performed to determine morphological cell death. DNA fragmentation and annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry were done to confirm apoptosis induced by SRJ23. Quantitation of cell cycle and apoptotic regulatory proteins were determined by immunoblotting.
Results
AGP and SRJ23 selectively inhibited the growth of prostate cancer cells compared with RAW 264.7 cells at low micromolar concentrations; however, SRJ23 was more potent. Mechanistically, SRJ23-treated PC-3 cells displayed down-regulation of cyclin-dependent kinase (CDK) 1 without affecting levels of CDK4 and cyclin D1. However, SRJ23 induced down-regulation of CDK4 and cyclin D1 but without affecting CDK1 in DU145 and LNCaP cell lines. DNA histogram analysis revealed that the SRJ23 induced G2/M in PC-3 cells but G1 arrest in DU-145 and LNCaP cells. Morphologically, both compounds induced predominantly apoptosis, which was further confirmed by DNA fragmentation and annexin V-FITC staining. The DNA fragmentation was inhibited in the presence of caspase 8 inhibitor (Z-IETD-FMK). Apoptosis was associated with an increase in caspase 8 expression and activation. This thought to have induced cleavage of Bid into t-Bid. Additionally, increased expression and activation of caspase 9 and Bax proteins were apparent, with a concomitant down-regulation of Bcl-2 protein. Similar apoptosis cascade of events was observed in SRJ23-treated DU145 and LNCaP cell lines.
Conclusion
SRJ23 inhibited the growth of prostate cancer cells by inducing G
2
/M and G1 arrest via down-regulation of CDK1, and CDK4 and cyclin, respectively, and initiated caspase-8-mediated mitochondrial apoptosis. Taken together, these data support the potential of this compound as a new anti-prostate cancer agent.</abstract><cop>Dordrecht</cop><pub>Springer Netherlands</pub><pmid>25070834</pmid><doi>10.1007/s10565-014-9282-5</doi><tpages>20</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0742-2091 |
| ispartof | Cell biology and toxicology, 2014-10, Vol.30 (5), p.269-288 |
| issn | 0742-2091 1573-6822 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_1618155351 |
| source | MEDLINE; SpringerNature Complete Journals |
| subjects | Animals Apoptosis Apoptosis - drug effects bcl-2-Associated X Protein - metabolism Biochemistry Biomedical and Life Sciences Caspase 8 - metabolism Caspase 9 - metabolism Cell Biology Cell cycle Cell Cycle Checkpoints - drug effects Cell Line, Tumor Cell Proliferation - drug effects Cyclin D1 - metabolism Cyclin-Dependent Kinases - metabolism Cytotoxicity Deoxyribonucleic acid Diterpenes - pharmacology DNA DNA Fragmentation - drug effects Down-Regulation - drug effects Fluorescence microscopy Guinea Pigs Humans Inhibitor drugs Iodides Life Sciences Male Original Research Pharmacology/Toxicology Prostate cancer Prostatic Neoplasms - drug therapy Prostatic Neoplasms - metabolism Proto-Oncogene Proteins c-bcl-2 - metabolism |
| title | SRJ23, a new semisynthetic andrographolide derivative: in vitro growth inhibition and mechanisms of cell cycle arrest and apoptosis in prostate cancer cells |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-28T05%3A30%3A33IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=SRJ23,%20a%20new%20semisynthetic%20andrographolide%20derivative:%20in%20vitro%20growth%20inhibition%20and%20mechanisms%20of%20cell%20cycle%20arrest%20and%20apoptosis%20in%20prostate%20cancer%20cells&rft.jtitle=Cell%20biology%20and%20toxicology&rft.au=Wong,%20Hui%20Chyn&rft.date=2014-10-01&rft.volume=30&rft.issue=5&rft.spage=269&rft.epage=288&rft.pages=269-288&rft.issn=0742-2091&rft.eissn=1573-6822&rft_id=info:doi/10.1007/s10565-014-9282-5&rft_dat=%3Cproquest_cross%3E3437560331%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1563758620&rft_id=info:pmid/25070834&rfr_iscdi=true |