RNA interference-mediated silencing of SOCS-1 via lentiviral vector promotes apoptosis of alveolar epithelial cells in vitro
Suppressor of cytokine signaling-1 (SOCS1) is a protein that negatively regulates cytokine and growth factor signaling. However, little is known regarding the precise role it plays in idiopathic pulmonary fibrosis. The aim of the present study was to construct a recombinant lentiviral vector for RNA...
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description | Suppressor of cytokine signaling-1 (SOCS1) is a protein that negatively regulates cytokine and growth factor signaling. However, little is known regarding the precise role it plays in idiopathic pulmonary fibrosis. The aim of the present study was to construct a recombinant lentiviral vector for RNA interference targeting the SOCS1 gene and to detect the expression in human alveolar epithelial cells. A lentiviral vector-mediated RNA interference method was used to establish a SOCS1-negative cell line of alveolar origin (A549). Three pairs of complementary small hairpin RNA (shRNA) oligonucleotides targeting the SOCS1 gene were designed, synthesized and inserted into the pPll3.7 vector. Packaged lentivirus particles were obtained after 48 h, and the supernatant was used to transfect the human alveolar epithelial cell line A549. The expression of the SOCS1 protein was detected by Western blotting. MTT assay was used to detect the cell proliferation of alveolar epithelial cells with SOCS1 knockdown. The recombinant plasmids were confirmed by sequencing. The lentivirus-containing supernatant effectively infected the A549 cell line, and the expression of SOCS1 protein was inhibited, which was confirmed by Western blotting in the target cells. MTT assay indicated the inhibition effect for cell proliferation of A549 cells in the SOCS1-RNA interference group, compared to the control group with no interference-mediated silencing of the SOCS1 gene. A lentiviral vector for RNA interference targeting the SOCS1 gene was successfully constructed, and cell survival tests showed that knockdown of the SOCS1 gene promotes the apoptosis of alveolar cells. |
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However, little is known regarding the precise role it plays in idiopathic pulmonary fibrosis. The aim of the present study was to construct a recombinant lentiviral vector for RNA interference targeting the SOCS1 gene and to detect the expression in human alveolar epithelial cells. A lentiviral vector-mediated RNA interference method was used to establish a SOCS1-negative cell line of alveolar origin (A549). Three pairs of complementary small hairpin RNA (shRNA) oligonucleotides targeting the SOCS1 gene were designed, synthesized and inserted into the pPll3.7 vector. Packaged lentivirus particles were obtained after 48 h, and the supernatant was used to transfect the human alveolar epithelial cell line A549. The expression of the SOCS1 protein was detected by Western blotting. MTT assay was used to detect the cell proliferation of alveolar epithelial cells with SOCS1 knockdown. The recombinant plasmids were confirmed by sequencing. The lentivirus-containing supernatant effectively infected the A549 cell line, and the expression of SOCS1 protein was inhibited, which was confirmed by Western blotting in the target cells. MTT assay indicated the inhibition effect for cell proliferation of A549 cells in the SOCS1-RNA interference group, compared to the control group with no interference-mediated silencing of the SOCS1 gene. A lentiviral vector for RNA interference targeting the SOCS1 gene was successfully constructed, and cell survival tests showed that knockdown of the SOCS1 gene promotes the apoptosis of alveolar cells.</description><identifier>ISSN: 1791-2997</identifier><identifier>EISSN: 1791-3004</identifier><identifier>DOI: 10.3892/mmr.2011.672</identifier><identifier>PMID: 22086204</identifier><language>eng</language><publisher>Greece: Spandidos Publications UK Ltd</publisher><subject>Adenocarcinoma, Bronchiolo-Alveolar - metabolism ; Adenocarcinoma, Bronchiolo-Alveolar - pathology ; Alveoli ; Apoptosis ; Base Sequence ; Cell growth ; Cell Line, Tumor ; Cell proliferation ; Cell survival ; Cellular biology ; Cytokines ; Epithelial cells ; Epithelial Cells - cytology ; Epithelial Cells - metabolism ; Fibrosis ; Gene Silencing ; Genetic Vectors - chemistry ; Genetic Vectors - metabolism ; Growth factors ; Humans ; Inflammation ; Kinases ; Lentivirus - genetics ; Lung diseases ; Molecular Sequence Data ; Oligonucleotides ; Plasmids ; Proteins ; Pulmonary fibrosis ; RNA Interference ; RNA, Small Interfering - metabolism ; RNA-mediated interference ; SOCS-1 protein ; Suppressor of Cytokine Signaling 1 Protein ; Suppressor of Cytokine Signaling Proteins - antagonists & inhibitors ; Suppressor of Cytokine Signaling Proteins - genetics ; Suppressor of Cytokine Signaling Proteins - metabolism ; Western blotting</subject><ispartof>Molecular medicine reports, 2012-02, Vol.5 (2), p.452-456</ispartof><rights>Copyright Spandidos Publications UK Ltd. 2012</rights><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c356t-bb30696499148d543570513b245fe46878be04bb0fe90977d29ce8c3f544e67a3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22086204$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Qian, Yan-Rong</creatorcontrib><creatorcontrib>Zhang, Qiu-Rui</creatorcontrib><creatorcontrib>Cheng, Ting</creatorcontrib><creatorcontrib>Wan, Huan-Ying</creatorcontrib><creatorcontrib>Zhou, Min</creatorcontrib><title>RNA interference-mediated silencing of SOCS-1 via lentiviral vector promotes apoptosis of alveolar epithelial cells in vitro</title><title>Molecular medicine reports</title><addtitle>Mol Med Rep</addtitle><description>Suppressor of cytokine signaling-1 (SOCS1) is a protein that negatively regulates cytokine and growth factor signaling. However, little is known regarding the precise role it plays in idiopathic pulmonary fibrosis. The aim of the present study was to construct a recombinant lentiviral vector for RNA interference targeting the SOCS1 gene and to detect the expression in human alveolar epithelial cells. A lentiviral vector-mediated RNA interference method was used to establish a SOCS1-negative cell line of alveolar origin (A549). Three pairs of complementary small hairpin RNA (shRNA) oligonucleotides targeting the SOCS1 gene were designed, synthesized and inserted into the pPll3.7 vector. Packaged lentivirus particles were obtained after 48 h, and the supernatant was used to transfect the human alveolar epithelial cell line A549. The expression of the SOCS1 protein was detected by Western blotting. MTT assay was used to detect the cell proliferation of alveolar epithelial cells with SOCS1 knockdown. The recombinant plasmids were confirmed by sequencing. The lentivirus-containing supernatant effectively infected the A549 cell line, and the expression of SOCS1 protein was inhibited, which was confirmed by Western blotting in the target cells. MTT assay indicated the inhibition effect for cell proliferation of A549 cells in the SOCS1-RNA interference group, compared to the control group with no interference-mediated silencing of the SOCS1 gene. A lentiviral vector for RNA interference targeting the SOCS1 gene was successfully constructed, and cell survival tests showed that knockdown of the SOCS1 gene promotes the apoptosis of alveolar cells.</description><subject>Adenocarcinoma, Bronchiolo-Alveolar - metabolism</subject><subject>Adenocarcinoma, Bronchiolo-Alveolar - pathology</subject><subject>Alveoli</subject><subject>Apoptosis</subject><subject>Base Sequence</subject><subject>Cell growth</subject><subject>Cell Line, Tumor</subject><subject>Cell proliferation</subject><subject>Cell survival</subject><subject>Cellular biology</subject><subject>Cytokines</subject><subject>Epithelial cells</subject><subject>Epithelial Cells - cytology</subject><subject>Epithelial Cells - metabolism</subject><subject>Fibrosis</subject><subject>Gene Silencing</subject><subject>Genetic Vectors - chemistry</subject><subject>Genetic Vectors - metabolism</subject><subject>Growth factors</subject><subject>Humans</subject><subject>Inflammation</subject><subject>Kinases</subject><subject>Lentivirus - genetics</subject><subject>Lung diseases</subject><subject>Molecular Sequence Data</subject><subject>Oligonucleotides</subject><subject>Plasmids</subject><subject>Proteins</subject><subject>Pulmonary fibrosis</subject><subject>RNA Interference</subject><subject>RNA, Small Interfering - metabolism</subject><subject>RNA-mediated interference</subject><subject>SOCS-1 protein</subject><subject>Suppressor of Cytokine Signaling 1 Protein</subject><subject>Suppressor of Cytokine Signaling Proteins - antagonists & inhibitors</subject><subject>Suppressor of Cytokine Signaling Proteins - genetics</subject><subject>Suppressor of Cytokine Signaling Proteins - metabolism</subject><subject>Western blotting</subject><issn>1791-2997</issn><issn>1791-3004</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNpdkc9LHDEYhoNUXF178yyBHnrprPmdyVEWWwvigtpzyMx-Y7PMTMYkuyD4x5vFbQ-eEj6e781LHoQuKFnw2rCrYYgLRihdKM2O0CnVhlacEPHlcGfG6Bk6S2lDiJJMmhM0Y4zUihFxit4e7q-xHzPEDiKMLVQDrL3LsMbJ92Xgx2ccOvy4Wj5WFO-8w2Wa_c5H1-MdtDlEPMUwhAwJuylMOSSf9iuu30HoXcQw-fwXel8WWuj7VN4rQTmGc3TcuT7B18M5R39-3jwtb6u71a_fy-u7quVS5appOFFGCWOoqNdScKmJpLxhQnYgVK3rBohoGtKBIUbrNTMt1C3vpBCgtONz9P0jtxR92ULKdvBpX8WNELbJGkqNoEqwQn77RG7CNo6lnGVaSUpqymmhfnxQbQwpRejsFP3g4qulxO6l2CLF7qXYIqXgl4fQbVN-9z_8zwJ_B_Rdh-c</recordid><startdate>20120201</startdate><enddate>20120201</enddate><creator>Qian, Yan-Rong</creator><creator>Zhang, Qiu-Rui</creator><creator>Cheng, Ting</creator><creator>Wan, Huan-Ying</creator><creator>Zhou, Min</creator><general>Spandidos Publications UK Ltd</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>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AN0</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope></search><sort><creationdate>20120201</creationdate><title>RNA interference-mediated silencing of SOCS-1 via lentiviral vector promotes apoptosis of alveolar epithelial cells in vitro</title><author>Qian, Yan-Rong ; Zhang, Qiu-Rui ; Cheng, Ting ; Wan, Huan-Ying ; Zhou, Min</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c356t-bb30696499148d543570513b245fe46878be04bb0fe90977d29ce8c3f544e67a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Adenocarcinoma, Bronchiolo-Alveolar - metabolism</topic><topic>Adenocarcinoma, Bronchiolo-Alveolar - pathology</topic><topic>Alveoli</topic><topic>Apoptosis</topic><topic>Base Sequence</topic><topic>Cell growth</topic><topic>Cell Line, Tumor</topic><topic>Cell proliferation</topic><topic>Cell survival</topic><topic>Cellular biology</topic><topic>Cytokines</topic><topic>Epithelial cells</topic><topic>Epithelial Cells - cytology</topic><topic>Epithelial Cells - metabolism</topic><topic>Fibrosis</topic><topic>Gene Silencing</topic><topic>Genetic Vectors - chemistry</topic><topic>Genetic Vectors - metabolism</topic><topic>Growth factors</topic><topic>Humans</topic><topic>Inflammation</topic><topic>Kinases</topic><topic>Lentivirus - genetics</topic><topic>Lung diseases</topic><topic>Molecular Sequence Data</topic><topic>Oligonucleotides</topic><topic>Plasmids</topic><topic>Proteins</topic><topic>Pulmonary fibrosis</topic><topic>RNA Interference</topic><topic>RNA, Small Interfering - metabolism</topic><topic>RNA-mediated interference</topic><topic>SOCS-1 protein</topic><topic>Suppressor of Cytokine Signaling 1 Protein</topic><topic>Suppressor of Cytokine Signaling Proteins - antagonists & inhibitors</topic><topic>Suppressor of Cytokine Signaling Proteins - genetics</topic><topic>Suppressor of Cytokine Signaling Proteins - metabolism</topic><topic>Western blotting</topic><toplevel>online_resources</toplevel><creatorcontrib>Qian, Yan-Rong</creatorcontrib><creatorcontrib>Zhang, Qiu-Rui</creatorcontrib><creatorcontrib>Cheng, Ting</creatorcontrib><creatorcontrib>Wan, Huan-Ying</creatorcontrib><creatorcontrib>Zhou, Min</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>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</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>British Nursing Database</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</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>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological 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>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><jtitle>Molecular medicine reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Qian, Yan-Rong</au><au>Zhang, Qiu-Rui</au><au>Cheng, Ting</au><au>Wan, Huan-Ying</au><au>Zhou, Min</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>RNA interference-mediated silencing of SOCS-1 via lentiviral vector promotes apoptosis of alveolar epithelial cells in vitro</atitle><jtitle>Molecular medicine reports</jtitle><addtitle>Mol Med Rep</addtitle><date>2012-02-01</date><risdate>2012</risdate><volume>5</volume><issue>2</issue><spage>452</spage><epage>456</epage><pages>452-456</pages><issn>1791-2997</issn><eissn>1791-3004</eissn><abstract>Suppressor of cytokine signaling-1 (SOCS1) is a protein that negatively regulates cytokine and growth factor signaling. However, little is known regarding the precise role it plays in idiopathic pulmonary fibrosis. The aim of the present study was to construct a recombinant lentiviral vector for RNA interference targeting the SOCS1 gene and to detect the expression in human alveolar epithelial cells. A lentiviral vector-mediated RNA interference method was used to establish a SOCS1-negative cell line of alveolar origin (A549). Three pairs of complementary small hairpin RNA (shRNA) oligonucleotides targeting the SOCS1 gene were designed, synthesized and inserted into the pPll3.7 vector. Packaged lentivirus particles were obtained after 48 h, and the supernatant was used to transfect the human alveolar epithelial cell line A549. The expression of the SOCS1 protein was detected by Western blotting. MTT assay was used to detect the cell proliferation of alveolar epithelial cells with SOCS1 knockdown. The recombinant plasmids were confirmed by sequencing. The lentivirus-containing supernatant effectively infected the A549 cell line, and the expression of SOCS1 protein was inhibited, which was confirmed by Western blotting in the target cells. MTT assay indicated the inhibition effect for cell proliferation of A549 cells in the SOCS1-RNA interference group, compared to the control group with no interference-mediated silencing of the SOCS1 gene. A lentiviral vector for RNA interference targeting the SOCS1 gene was successfully constructed, and cell survival tests showed that knockdown of the SOCS1 gene promotes the apoptosis of alveolar cells.</abstract><cop>Greece</cop><pub>Spandidos Publications UK Ltd</pub><pmid>22086204</pmid><doi>10.3892/mmr.2011.672</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Adenocarcinoma, Bronchiolo-Alveolar - metabolism Adenocarcinoma, Bronchiolo-Alveolar - pathology Alveoli Apoptosis Base Sequence Cell growth Cell Line, Tumor Cell proliferation Cell survival Cellular biology Cytokines Epithelial cells Epithelial Cells - cytology Epithelial Cells - metabolism Fibrosis Gene Silencing Genetic Vectors - chemistry Genetic Vectors - metabolism Growth factors Humans Inflammation Kinases Lentivirus - genetics Lung diseases Molecular Sequence Data Oligonucleotides Plasmids Proteins Pulmonary fibrosis RNA Interference RNA, Small Interfering - metabolism RNA-mediated interference SOCS-1 protein Suppressor of Cytokine Signaling 1 Protein Suppressor of Cytokine Signaling Proteins - antagonists & inhibitors Suppressor of Cytokine Signaling Proteins - genetics Suppressor of Cytokine Signaling Proteins - metabolism Western blotting |
title | RNA interference-mediated silencing of SOCS-1 via lentiviral vector promotes apoptosis of alveolar epithelial cells in vitro |
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