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Clusterin silencing inhibits proliferation and reduces invasion in human laryngeal squamous carcinoma cells
Clusterin is, in its major form, a secreted heterodimeric disulfide-linked glycoprotein (sCLU), which plays important roles in cell survival and death. In laryngeal squamous cell carcinomas (LSCC), sCLU is up-regulated and its expression is related to the invasiveness of these tumors. The purpose of...
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| Published in: | World journal of surgical oncology 2014-04, Vol.12 (1), p.124-124, Article 124 |
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| creator | Wang, Qianjin Cao, Weiyan Su, Quancai Liu, Zimin Zhang, Lin |
| description | Clusterin is, in its major form, a secreted heterodimeric disulfide-linked glycoprotein (sCLU), which plays important roles in cell survival and death. In laryngeal squamous cell carcinomas (LSCC), sCLU is up-regulated and its expression is related to the invasiveness of these tumors. The purpose of this study was to explore the inhibiting role of sCLU gene silence in the invasive ability and growth of Hep-2 human laryngeal squamous carcinoma cells (Hep-2) by transfection of short hairpin RNA expression plasmids against sCLU (sCLU-shRNA) (in vivo) or small interference RNA (sCLU-siRNA) (in vitro).
sCLU-siRNA and the control siRNA were transfected into Hep-2 cells using Lipofectamine 2000. RT-PCR and Western blot were used to detect the effect of siRNA transfection on sCLU mRNA and sCLU protein expression. The invasive activity of sCLU-siRNA-transfected Hep-2 cells was measured with the modified Boyden chamber assay and wound healing assay. The effects of sCLU-siRNA on cell proliferation were evaluated by MTT assay. Apoptosis was measured by Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double-staining methods. We next evaluated the effects of sCLU silencing by sCLU-shRNA transfection in vivo on tumor growth and metastatic properties to the lung. Terminal deoxytransferase-mediated dUTP nick end labeling (TUNEL) staining was used to observe the apoptosis in the xenografts.
It showed that siRNA-mediated down-regulation of sCLU expression in Hep-2 cells significantly inhibited cell proliferation and promoted apoptosis in vitro. Furthermore, siRNA-mediated down-regulation of sCLU expression decreases in vitro cell migration and invasion ability. In vivo, the average volume of tumors in the sCLU-shRNA transfected group was significantly lower than in the control group (P |
| doi_str_mv | 10.1186/1477-7819-12-124 |
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sCLU-siRNA and the control siRNA were transfected into Hep-2 cells using Lipofectamine 2000. RT-PCR and Western blot were used to detect the effect of siRNA transfection on sCLU mRNA and sCLU protein expression. The invasive activity of sCLU-siRNA-transfected Hep-2 cells was measured with the modified Boyden chamber assay and wound healing assay. The effects of sCLU-siRNA on cell proliferation were evaluated by MTT assay. Apoptosis was measured by Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double-staining methods. We next evaluated the effects of sCLU silencing by sCLU-shRNA transfection in vivo on tumor growth and metastatic properties to the lung. Terminal deoxytransferase-mediated dUTP nick end labeling (TUNEL) staining was used to observe the apoptosis in the xenografts.
It showed that siRNA-mediated down-regulation of sCLU expression in Hep-2 cells significantly inhibited cell proliferation and promoted apoptosis in vitro. Furthermore, siRNA-mediated down-regulation of sCLU expression decreases in vitro cell migration and invasion ability. In vivo, the average volume of tumors in the sCLU-shRNA transfected group was significantly lower than in the control group (P<0.01), and the significant apoptosis detected with TUNEL was indicated in the sCLU-shRNA transfected groups (P<0.05). Significantly, we found that sCLU-shRNA could exert marked inhibition of the lung metastasis of Hep-2 cells in nude mice in vivo.
sCLU gene silence can inhibit invasion and growth of LSCC. sCLU may provide a potential therapeutic target against human LSCC.</description><identifier>ISSN: 1477-7819</identifier><identifier>EISSN: 1477-7819</identifier><identifier>DOI: 10.1186/1477-7819-12-124</identifier><identifier>PMID: 24767179</identifier><language>eng</language><publisher>England: BioMed Central Ltd</publisher><subject>Analysis ; Animals ; Apoptosis ; Blotting, Western ; Breast cancer ; Carcinoma, Squamous Cell - metabolism ; Carcinoma, Squamous Cell - pathology ; Carcinoma, Squamous Cell - prevention & control ; Cell Adhesion ; Cell culture ; Cell Movement ; Cell Proliferation ; Clusterin - antagonists & inhibitors ; Clusterin - genetics ; Clusterin - metabolism ; Data analysis ; Female ; Gene expression ; Gene Silencing ; Humans ; Laryngeal cancer ; Laryngeal Neoplasms - metabolism ; Laryngeal Neoplasms - pathology ; Laryngeal Neoplasms - prevention & control ; Lung Neoplasms - metabolism ; Lung Neoplasms - prevention & control ; Lung Neoplasms - secondary ; Membranes ; Metastasis ; Mice ; Mice, SCID ; Neoplasm Invasiveness ; Physiological aspects ; Plasmids ; Proteins ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger - genetics ; RNA, Small Interfering - genetics ; Squamous cell carcinoma ; Tumor Cells, Cultured ; Tumors ; Wound Healing</subject><ispartof>World journal of surgical oncology, 2014-04, Vol.12 (1), p.124-124, Article 124</ispartof><rights>COPYRIGHT 2014 BioMed Central Ltd.</rights><rights>2014 Wang et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.</rights><rights>Copyright © 2014 Wang et al.; licensee BioMed Central Ltd. 2014 Wang et al.; licensee BioMed Central Ltd.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-b648t-54b710739b9adc49fa8cec2893da888fd85a1317bdd2de6b5432a2edcf80ab233</citedby><cites>FETCH-LOGICAL-b648t-54b710739b9adc49fa8cec2893da888fd85a1317bdd2de6b5432a2edcf80ab233</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4016627/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/1522968992?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,25753,27924,27925,37012,37013,44590,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24767179$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wang, Qianjin</creatorcontrib><creatorcontrib>Cao, Weiyan</creatorcontrib><creatorcontrib>Su, Quancai</creatorcontrib><creatorcontrib>Liu, Zimin</creatorcontrib><creatorcontrib>Zhang, Lin</creatorcontrib><title>Clusterin silencing inhibits proliferation and reduces invasion in human laryngeal squamous carcinoma cells</title><title>World journal of surgical oncology</title><addtitle>World J Surg Oncol</addtitle><description>Clusterin is, in its major form, a secreted heterodimeric disulfide-linked glycoprotein (sCLU), which plays important roles in cell survival and death. In laryngeal squamous cell carcinomas (LSCC), sCLU is up-regulated and its expression is related to the invasiveness of these tumors. The purpose of this study was to explore the inhibiting role of sCLU gene silence in the invasive ability and growth of Hep-2 human laryngeal squamous carcinoma cells (Hep-2) by transfection of short hairpin RNA expression plasmids against sCLU (sCLU-shRNA) (in vivo) or small interference RNA (sCLU-siRNA) (in vitro).
sCLU-siRNA and the control siRNA were transfected into Hep-2 cells using Lipofectamine 2000. RT-PCR and Western blot were used to detect the effect of siRNA transfection on sCLU mRNA and sCLU protein expression. The invasive activity of sCLU-siRNA-transfected Hep-2 cells was measured with the modified Boyden chamber assay and wound healing assay. The effects of sCLU-siRNA on cell proliferation were evaluated by MTT assay. Apoptosis was measured by Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double-staining methods. We next evaluated the effects of sCLU silencing by sCLU-shRNA transfection in vivo on tumor growth and metastatic properties to the lung. Terminal deoxytransferase-mediated dUTP nick end labeling (TUNEL) staining was used to observe the apoptosis in the xenografts.
It showed that siRNA-mediated down-regulation of sCLU expression in Hep-2 cells significantly inhibited cell proliferation and promoted apoptosis in vitro. Furthermore, siRNA-mediated down-regulation of sCLU expression decreases in vitro cell migration and invasion ability. In vivo, the average volume of tumors in the sCLU-shRNA transfected group was significantly lower than in the control group (P<0.01), and the significant apoptosis detected with TUNEL was indicated in the sCLU-shRNA transfected groups (P<0.05). Significantly, we found that sCLU-shRNA could exert marked inhibition of the lung metastasis of Hep-2 cells in nude mice in vivo.
sCLU gene silence can inhibit invasion and growth of LSCC. sCLU may provide a potential therapeutic target against human LSCC.</description><subject>Analysis</subject><subject>Animals</subject><subject>Apoptosis</subject><subject>Blotting, Western</subject><subject>Breast cancer</subject><subject>Carcinoma, Squamous Cell - metabolism</subject><subject>Carcinoma, Squamous Cell - pathology</subject><subject>Carcinoma, Squamous Cell - prevention & control</subject><subject>Cell Adhesion</subject><subject>Cell culture</subject><subject>Cell Movement</subject><subject>Cell Proliferation</subject><subject>Clusterin - antagonists & inhibitors</subject><subject>Clusterin - genetics</subject><subject>Clusterin - metabolism</subject><subject>Data analysis</subject><subject>Female</subject><subject>Gene expression</subject><subject>Gene Silencing</subject><subject>Humans</subject><subject>Laryngeal cancer</subject><subject>Laryngeal Neoplasms - metabolism</subject><subject>Laryngeal Neoplasms - pathology</subject><subject>Laryngeal Neoplasms - prevention & control</subject><subject>Lung Neoplasms - metabolism</subject><subject>Lung Neoplasms - prevention & control</subject><subject>Lung Neoplasms - secondary</subject><subject>Membranes</subject><subject>Metastasis</subject><subject>Mice</subject><subject>Mice, SCID</subject><subject>Neoplasm Invasiveness</subject><subject>Physiological aspects</subject><subject>Plasmids</subject><subject>Proteins</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Small Interfering - genetics</subject><subject>Squamous cell carcinoma</subject><subject>Tumor Cells, Cultured</subject><subject>Tumors</subject><subject>Wound Healing</subject><issn>1477-7819</issn><issn>1477-7819</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>PIMPY</sourceid><recordid>eNp1kt2L1DAUxYso7rr67pMUBPGla5O0-XgRlsEvWPBFn8Ntmk6zpsls0i7sf-8ts44zspJAys3vnibnpChek_qSEMk_kEaISkiiKkJxNk-K80Pp6dH3WfEi55u6poy17HlxRhvBBRHqvPi18UuebXKhzM7bYFzYli6MrnNzLncpejfYBLOLoYTQl8n2i7EZkTvIaxEbx2WCUHpI92FrwZf5doEpLrk0kFAvTlAa631-WTwbwGf76mG9KH5-_vRj87W6_v7l2-bquup4I-eqbTpBasFUp6A3jRpAGmuoVKwHKeXQyxYII6Lre9pb3rUNo0BtbwZZQ4dXvCg-7nV3Szdh3YY5gde75CY8o47g9OlOcKPexjvd1IRzKlBgsxfoXPyPwOmOiZNezdar2ZpQnA2qvH84Roq3i82znlxejYBg0R1NWoY54I04om__QW_ikgKahBSlikul6F9qC95qF4aIPzerqL5qmeKK81YhdfkIhaO3kzMx2AFzPm14d9QwYoLzmKNf1szzKVjvQZNizskOB0dIrdf3-JgHb46jODT8eYDsNxKv3PU</recordid><startdate>20140426</startdate><enddate>20140426</enddate><creator>Wang, Qianjin</creator><creator>Cao, Weiyan</creator><creator>Su, Quancai</creator><creator>Liu, Zimin</creator><creator>Zhang, Lin</creator><general>BioMed Central Ltd</general><general>BioMed Central</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>7QL</scope><scope>7QO</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FD</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>P64</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20140426</creationdate><title>Clusterin silencing inhibits proliferation and reduces invasion in human laryngeal squamous carcinoma cells</title><author>Wang, Qianjin ; Cao, Weiyan ; Su, Quancai ; Liu, Zimin ; Zhang, Lin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-b648t-54b710739b9adc49fa8cec2893da888fd85a1317bdd2de6b5432a2edcf80ab233</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Analysis</topic><topic>Animals</topic><topic>Apoptosis</topic><topic>Blotting, Western</topic><topic>Breast cancer</topic><topic>Carcinoma, Squamous Cell - metabolism</topic><topic>Carcinoma, Squamous Cell - pathology</topic><topic>Carcinoma, Squamous Cell - prevention & control</topic><topic>Cell Adhesion</topic><topic>Cell culture</topic><topic>Cell Movement</topic><topic>Cell Proliferation</topic><topic>Clusterin - antagonists & inhibitors</topic><topic>Clusterin - genetics</topic><topic>Clusterin - metabolism</topic><topic>Data analysis</topic><topic>Female</topic><topic>Gene expression</topic><topic>Gene Silencing</topic><topic>Humans</topic><topic>Laryngeal cancer</topic><topic>Laryngeal Neoplasms - metabolism</topic><topic>Laryngeal Neoplasms - pathology</topic><topic>Laryngeal Neoplasms - prevention & control</topic><topic>Lung Neoplasms - metabolism</topic><topic>Lung Neoplasms - prevention & control</topic><topic>Lung Neoplasms - secondary</topic><topic>Membranes</topic><topic>Metastasis</topic><topic>Mice</topic><topic>Mice, SCID</topic><topic>Neoplasm Invasiveness</topic><topic>Physiological aspects</topic><topic>Plasmids</topic><topic>Proteins</topic><topic>Real-Time Polymerase Chain Reaction</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Small Interfering - genetics</topic><topic>Squamous cell carcinoma</topic><topic>Tumor Cells, Cultured</topic><topic>Tumors</topic><topic>Wound Healing</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Qianjin</creatorcontrib><creatorcontrib>Cao, Weiyan</creatorcontrib><creatorcontrib>Su, Quancai</creatorcontrib><creatorcontrib>Liu, Zimin</creatorcontrib><creatorcontrib>Zhang, Lin</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>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Technology Research Database</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>ProQuest Central Essentials</collection><collection>ProQuest Central</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 Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Publicly Available Content 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><collection>PubMed Central (Full Participant titles)</collection><jtitle>World journal of surgical oncology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Qianjin</au><au>Cao, Weiyan</au><au>Su, Quancai</au><au>Liu, Zimin</au><au>Zhang, Lin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Clusterin silencing inhibits proliferation and reduces invasion in human laryngeal squamous carcinoma cells</atitle><jtitle>World journal of surgical oncology</jtitle><addtitle>World J Surg Oncol</addtitle><date>2014-04-26</date><risdate>2014</risdate><volume>12</volume><issue>1</issue><spage>124</spage><epage>124</epage><pages>124-124</pages><artnum>124</artnum><issn>1477-7819</issn><eissn>1477-7819</eissn><abstract>Clusterin is, in its major form, a secreted heterodimeric disulfide-linked glycoprotein (sCLU), which plays important roles in cell survival and death. In laryngeal squamous cell carcinomas (LSCC), sCLU is up-regulated and its expression is related to the invasiveness of these tumors. The purpose of this study was to explore the inhibiting role of sCLU gene silence in the invasive ability and growth of Hep-2 human laryngeal squamous carcinoma cells (Hep-2) by transfection of short hairpin RNA expression plasmids against sCLU (sCLU-shRNA) (in vivo) or small interference RNA (sCLU-siRNA) (in vitro).
sCLU-siRNA and the control siRNA were transfected into Hep-2 cells using Lipofectamine 2000. RT-PCR and Western blot were used to detect the effect of siRNA transfection on sCLU mRNA and sCLU protein expression. The invasive activity of sCLU-siRNA-transfected Hep-2 cells was measured with the modified Boyden chamber assay and wound healing assay. The effects of sCLU-siRNA on cell proliferation were evaluated by MTT assay. Apoptosis was measured by Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double-staining methods. We next evaluated the effects of sCLU silencing by sCLU-shRNA transfection in vivo on tumor growth and metastatic properties to the lung. Terminal deoxytransferase-mediated dUTP nick end labeling (TUNEL) staining was used to observe the apoptosis in the xenografts.
It showed that siRNA-mediated down-regulation of sCLU expression in Hep-2 cells significantly inhibited cell proliferation and promoted apoptosis in vitro. Furthermore, siRNA-mediated down-regulation of sCLU expression decreases in vitro cell migration and invasion ability. In vivo, the average volume of tumors in the sCLU-shRNA transfected group was significantly lower than in the control group (P<0.01), and the significant apoptosis detected with TUNEL was indicated in the sCLU-shRNA transfected groups (P<0.05). Significantly, we found that sCLU-shRNA could exert marked inhibition of the lung metastasis of Hep-2 cells in nude mice in vivo.
sCLU gene silence can inhibit invasion and growth of LSCC. sCLU may provide a potential therapeutic target against human LSCC.</abstract><cop>England</cop><pub>BioMed Central Ltd</pub><pmid>24767179</pmid><doi>10.1186/1477-7819-12-124</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Analysis Animals Apoptosis Blotting, Western Breast cancer Carcinoma, Squamous Cell - metabolism Carcinoma, Squamous Cell - pathology Carcinoma, Squamous Cell - prevention & control Cell Adhesion Cell culture Cell Movement Cell Proliferation Clusterin - antagonists & inhibitors Clusterin - genetics Clusterin - metabolism Data analysis Female Gene expression Gene Silencing Humans Laryngeal cancer Laryngeal Neoplasms - metabolism Laryngeal Neoplasms - pathology Laryngeal Neoplasms - prevention & control Lung Neoplasms - metabolism Lung Neoplasms - prevention & control Lung Neoplasms - secondary Membranes Metastasis Mice Mice, SCID Neoplasm Invasiveness Physiological aspects Plasmids Proteins Real-Time Polymerase Chain Reaction Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger - genetics RNA, Small Interfering - genetics Squamous cell carcinoma Tumor Cells, Cultured Tumors Wound Healing |
| title | Clusterin silencing inhibits proliferation and reduces invasion in human laryngeal squamous carcinoma cells |
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