Suppression of tumorigenesis and metastasis of hepatocellular carcinoma by shRNA interference targeting on homeoprotein Six1
We previously demonstrated that the overexpression of homeoprotein Six1 in hepatocellular carcinoma (HCC) patients is associated with venous infiltration, advanced pathologic tumor metastasis (pTNM) stage and poor overall survival rate (Ng et al. Br J Cancer 2006;95:1050–5). In this study, short hai...
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| Veröffentlicht in: | International journal of cancer 2010-08, Vol.127 (4), p.859-872 |
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| creator | Ng, Kevin T.P. Lee, Terence K.W. Cheng, Qiao Wo, Jana Y.H. Sun, Chris K.W. Guo, Dong‐Yong Lim, Zophia X. Lo, Chung‐Mau Poon, Ronnie T.P. Fan, Sheung‐Tat Man, Kwan |
| description | We previously demonstrated that the overexpression of homeoprotein Six1 in hepatocellular carcinoma (HCC) patients is associated with venous infiltration, advanced pathologic tumor metastasis (pTNM) stage and poor overall survival rate (Ng et al. Br J Cancer 2006;95:1050–5). In this study, short hairpin RNA (shRNA) interference approach was used to suppress the expression of Six1 in a metastatic HCC cell line MHCC97L. Stable transfectant MHCC97L‐shSix1 carrying Six1‐specific shRNA plasmid was established to downregulate Six1 expression to about 40% when compared with MHCC97L‐Control. In vitro functional assays demonstrated that the growth rate and proliferation ability of MHCC97L‐shSix1 cells were markedly decreased. Moreover, significant decrease of cell motility and invasiveness were observed in MHCC97L‐shSix1 cells. Data from in vivo xenograft tumorigenesis model demonstrated that the size of tumor in MHCC97L‐shSix1 group was dramatically reduced. Experimental and spontaneous metastasis models indicated that targeting Six1 suppression noticeably reduced the pulmonary metastasis in MHCC97L‐shSix1 group. To identify Six1‐regulated targets, cDNA microarray was employed to compare the expression profiles of MHCC97L‐Control and MHCC97L‐shSix1 cells. Twenty‐eight downregulated and 24 upregulated genes with known functions were identified in MHCC97L‐shSix1. The functions of these target genes are involved in diverse biological activities. Our data suggest that Six1 may be involved in regulation of proliferation and invasiveness of HCC; thus targeting suppression of Six1 is a viable option for treating HCC patients. |
| doi_str_mv | 10.1002/ijc.25105 |
| format | Article |
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Br J Cancer 2006;95:1050–5). In this study, short hairpin RNA (shRNA) interference approach was used to suppress the expression of Six1 in a metastatic HCC cell line MHCC97L. Stable transfectant MHCC97L‐shSix1 carrying Six1‐specific shRNA plasmid was established to downregulate Six1 expression to about 40% when compared with MHCC97L‐Control. In vitro functional assays demonstrated that the growth rate and proliferation ability of MHCC97L‐shSix1 cells were markedly decreased. Moreover, significant decrease of cell motility and invasiveness were observed in MHCC97L‐shSix1 cells. Data from in vivo xenograft tumorigenesis model demonstrated that the size of tumor in MHCC97L‐shSix1 group was dramatically reduced. Experimental and spontaneous metastasis models indicated that targeting Six1 suppression noticeably reduced the pulmonary metastasis in MHCC97L‐shSix1 group. To identify Six1‐regulated targets, cDNA microarray was employed to compare the expression profiles of MHCC97L‐Control and MHCC97L‐shSix1 cells. Twenty‐eight downregulated and 24 upregulated genes with known functions were identified in MHCC97L‐shSix1. The functions of these target genes are involved in diverse biological activities. Our data suggest that Six1 may be involved in regulation of proliferation and invasiveness of HCC; thus targeting suppression of Six1 is a viable option for treating HCC patients.</description><identifier>ISSN: 0020-7136</identifier><identifier>EISSN: 1097-0215</identifier><identifier>DOI: 10.1002/ijc.25105</identifier><identifier>PMID: 20013809</identifier><identifier>CODEN: IJCNAW</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Animals ; Biological and medical sciences ; Biomarkers, Tumor - genetics ; Biomarkers, Tumor - metabolism ; Blotting, Western ; Carcinoma, Hepatocellular - genetics ; Carcinoma, Hepatocellular - prevention & control ; Carcinoma, Hepatocellular - secondary ; cDNA microarray ; Cell Adhesion ; Cell Cycle ; Cell Movement ; Cell Proliferation ; Colony-Forming Units Assay ; Fluorescent Antibody Technique ; Gastroenterology. Liver. Pancreas. Abdomen ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; hepatocellular carcinoma ; Homeodomain Proteins - antagonists & inhibitors ; Homeodomain Proteins - genetics ; Homeodomain Proteins - metabolism ; homeoprotein Six1 ; Humans ; interference ; Kidney Neoplasms - genetics ; Kidney Neoplasms - prevention & control ; Kidney Neoplasms - secondary ; Liver Neoplasms, Experimental - genetics ; Liver Neoplasms, Experimental - pathology ; Liver Neoplasms, Experimental - prevention & control ; Liver. Biliary tract. Portal circulation. Exocrine pancreas ; Medical sciences ; metastasis ; Mice ; Mice, Nude ; Oligonucleotide Array Sequence Analysis ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; RNA, Small Interfering - pharmacology ; short hairpin RNA (shRNA) ; Splenic Neoplasms - genetics ; Splenic Neoplasms - prevention & control ; Splenic Neoplasms - secondary ; Tumors ; Wound Healing</subject><ispartof>International journal of cancer, 2010-08, Vol.127 (4), p.859-872</ispartof><rights>Copyright © 2009 UICC</rights><rights>2015 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4555-86b56450ed9140f18927652b5959eea58c23307a717fcb409fe35eb583cbdee13</citedby><cites>FETCH-LOGICAL-c4555-86b56450ed9140f18927652b5959eea58c23307a717fcb409fe35eb583cbdee13</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fijc.25105$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fijc.25105$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27903,27904,45553,45554</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=22956196$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20013809$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ng, Kevin T.P.</creatorcontrib><creatorcontrib>Lee, Terence K.W.</creatorcontrib><creatorcontrib>Cheng, Qiao</creatorcontrib><creatorcontrib>Wo, Jana Y.H.</creatorcontrib><creatorcontrib>Sun, Chris K.W.</creatorcontrib><creatorcontrib>Guo, Dong‐Yong</creatorcontrib><creatorcontrib>Lim, Zophia X.</creatorcontrib><creatorcontrib>Lo, Chung‐Mau</creatorcontrib><creatorcontrib>Poon, Ronnie T.P.</creatorcontrib><creatorcontrib>Fan, Sheung‐Tat</creatorcontrib><creatorcontrib>Man, Kwan</creatorcontrib><title>Suppression of tumorigenesis and metastasis of hepatocellular carcinoma by shRNA interference targeting on homeoprotein Six1</title><title>International journal of cancer</title><addtitle>Int J Cancer</addtitle><description>We previously demonstrated that the overexpression of homeoprotein Six1 in hepatocellular carcinoma (HCC) patients is associated with venous infiltration, advanced pathologic tumor metastasis (pTNM) stage and poor overall survival rate (Ng et al. Br J Cancer 2006;95:1050–5). In this study, short hairpin RNA (shRNA) interference approach was used to suppress the expression of Six1 in a metastatic HCC cell line MHCC97L. Stable transfectant MHCC97L‐shSix1 carrying Six1‐specific shRNA plasmid was established to downregulate Six1 expression to about 40% when compared with MHCC97L‐Control. In vitro functional assays demonstrated that the growth rate and proliferation ability of MHCC97L‐shSix1 cells were markedly decreased. Moreover, significant decrease of cell motility and invasiveness were observed in MHCC97L‐shSix1 cells. Data from in vivo xenograft tumorigenesis model demonstrated that the size of tumor in MHCC97L‐shSix1 group was dramatically reduced. Experimental and spontaneous metastasis models indicated that targeting Six1 suppression noticeably reduced the pulmonary metastasis in MHCC97L‐shSix1 group. To identify Six1‐regulated targets, cDNA microarray was employed to compare the expression profiles of MHCC97L‐Control and MHCC97L‐shSix1 cells. Twenty‐eight downregulated and 24 upregulated genes with known functions were identified in MHCC97L‐shSix1. The functions of these target genes are involved in diverse biological activities. Our data suggest that Six1 may be involved in regulation of proliferation and invasiveness of HCC; thus targeting suppression of Six1 is a viable option for treating HCC patients.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Biomarkers, Tumor - genetics</subject><subject>Biomarkers, Tumor - metabolism</subject><subject>Blotting, Western</subject><subject>Carcinoma, Hepatocellular - genetics</subject><subject>Carcinoma, Hepatocellular - prevention & control</subject><subject>Carcinoma, Hepatocellular - secondary</subject><subject>cDNA microarray</subject><subject>Cell Adhesion</subject><subject>Cell Cycle</subject><subject>Cell Movement</subject><subject>Cell Proliferation</subject><subject>Colony-Forming Units Assay</subject><subject>Fluorescent Antibody Technique</subject><subject>Gastroenterology. Liver. Pancreas. Abdomen</subject><subject>Gene Expression Profiling</subject><subject>Gene Expression Regulation, Neoplastic</subject><subject>hepatocellular carcinoma</subject><subject>Homeodomain Proteins - antagonists & inhibitors</subject><subject>Homeodomain Proteins - genetics</subject><subject>Homeodomain Proteins - metabolism</subject><subject>homeoprotein Six1</subject><subject>Humans</subject><subject>interference</subject><subject>Kidney Neoplasms - genetics</subject><subject>Kidney Neoplasms - prevention & control</subject><subject>Kidney Neoplasms - secondary</subject><subject>Liver Neoplasms, Experimental - genetics</subject><subject>Liver Neoplasms, Experimental - pathology</subject><subject>Liver Neoplasms, Experimental - prevention & control</subject><subject>Liver. Biliary tract. Portal circulation. Exocrine pancreas</subject><subject>Medical sciences</subject><subject>metastasis</subject><subject>Mice</subject><subject>Mice, Nude</subject><subject>Oligonucleotide Array Sequence Analysis</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>RNA, Small Interfering - pharmacology</subject><subject>short hairpin RNA (shRNA)</subject><subject>Splenic Neoplasms - genetics</subject><subject>Splenic Neoplasms - prevention & control</subject><subject>Splenic Neoplasms - secondary</subject><subject>Tumors</subject><subject>Wound Healing</subject><issn>0020-7136</issn><issn>1097-0215</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kNFr2zAQxsVoadOsD_0Hil7G6IMTyfLZ1mMIbdcRNli2ZyMr50TFljzJpgvsj6-6ZO1T4eDuuB_fx32EXHE244ylc_OoZylwBh_IhDNZJCzlcEIm8caSgov8nFyE8MgY58CyM3KexlGUTE7I3_XY9x5DMM5S19Bh7Jw3W7QYTKDKbmiHgwqx4hrvO-zV4DS27dgqT7Xy2ljXKVrvadj9-Lagxg7oG_RoNdJB-S0Oxm5plN-5Dl3v3YDG0rX5wz-S00a1AS-PfUp-3d3-XH5JVt_vH5aLVaIzAEjKvIY8A4YbyTPW8FKmRQ5pDRIkooJSp0KwQhW8aHSdMdmgAKyhFLreIHIxJZ8PutH894hhqDoTXn5QFt0YqkKIDAoAEcmbA6m9C8FjU_XedMrvK86ql6yrmHX1L-vIXh9Vx7rDzSv5P9wIfDoCKmjVNl5ZbcIbl0rIucwjNz9wT6bF_fuO1cPX5cH6Gdxdlyk</recordid><startdate>20100815</startdate><enddate>20100815</enddate><creator>Ng, Kevin T.P.</creator><creator>Lee, Terence K.W.</creator><creator>Cheng, Qiao</creator><creator>Wo, Jana Y.H.</creator><creator>Sun, Chris K.W.</creator><creator>Guo, Dong‐Yong</creator><creator>Lim, Zophia X.</creator><creator>Lo, Chung‐Mau</creator><creator>Poon, Ronnie T.P.</creator><creator>Fan, Sheung‐Tat</creator><creator>Man, Kwan</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><general>Wiley-Blackwell</general><scope>IQODW</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>20100815</creationdate><title>Suppression of tumorigenesis and metastasis of hepatocellular carcinoma by shRNA interference targeting on homeoprotein Six1</title><author>Ng, Kevin T.P. ; Lee, Terence K.W. ; Cheng, Qiao ; Wo, Jana Y.H. ; Sun, Chris K.W. ; Guo, Dong‐Yong ; Lim, Zophia X. ; Lo, Chung‐Mau ; Poon, Ronnie T.P. ; Fan, Sheung‐Tat ; Man, Kwan</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4555-86b56450ed9140f18927652b5959eea58c23307a717fcb409fe35eb583cbdee13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Biomarkers, Tumor - genetics</topic><topic>Biomarkers, Tumor - metabolism</topic><topic>Blotting, Western</topic><topic>Carcinoma, Hepatocellular - genetics</topic><topic>Carcinoma, Hepatocellular - prevention & control</topic><topic>Carcinoma, Hepatocellular - secondary</topic><topic>cDNA microarray</topic><topic>Cell Adhesion</topic><topic>Cell Cycle</topic><topic>Cell Movement</topic><topic>Cell Proliferation</topic><topic>Colony-Forming Units Assay</topic><topic>Fluorescent Antibody Technique</topic><topic>Gastroenterology. Liver. Pancreas. Abdomen</topic><topic>Gene Expression Profiling</topic><topic>Gene Expression Regulation, Neoplastic</topic><topic>hepatocellular carcinoma</topic><topic>Homeodomain Proteins - antagonists & inhibitors</topic><topic>Homeodomain Proteins - genetics</topic><topic>Homeodomain Proteins - metabolism</topic><topic>homeoprotein Six1</topic><topic>Humans</topic><topic>interference</topic><topic>Kidney Neoplasms - genetics</topic><topic>Kidney Neoplasms - prevention & control</topic><topic>Kidney Neoplasms - secondary</topic><topic>Liver Neoplasms, Experimental - genetics</topic><topic>Liver Neoplasms, Experimental - pathology</topic><topic>Liver Neoplasms, Experimental - prevention & control</topic><topic>Liver. Biliary tract. Portal circulation. Exocrine pancreas</topic><topic>Medical sciences</topic><topic>metastasis</topic><topic>Mice</topic><topic>Mice, Nude</topic><topic>Oligonucleotide Array Sequence Analysis</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Messenger - metabolism</topic><topic>RNA, Small Interfering - pharmacology</topic><topic>short hairpin RNA (shRNA)</topic><topic>Splenic Neoplasms - genetics</topic><topic>Splenic Neoplasms - prevention & control</topic><topic>Splenic Neoplasms - secondary</topic><topic>Tumors</topic><topic>Wound Healing</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ng, Kevin T.P.</creatorcontrib><creatorcontrib>Lee, Terence K.W.</creatorcontrib><creatorcontrib>Cheng, Qiao</creatorcontrib><creatorcontrib>Wo, Jana Y.H.</creatorcontrib><creatorcontrib>Sun, Chris K.W.</creatorcontrib><creatorcontrib>Guo, Dong‐Yong</creatorcontrib><creatorcontrib>Lim, Zophia X.</creatorcontrib><creatorcontrib>Lo, Chung‐Mau</creatorcontrib><creatorcontrib>Poon, Ronnie T.P.</creatorcontrib><creatorcontrib>Fan, Sheung‐Tat</creatorcontrib><creatorcontrib>Man, Kwan</creatorcontrib><collection>Pascal-Francis</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>International journal of cancer</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ng, Kevin T.P.</au><au>Lee, Terence K.W.</au><au>Cheng, Qiao</au><au>Wo, Jana Y.H.</au><au>Sun, Chris K.W.</au><au>Guo, Dong‐Yong</au><au>Lim, Zophia X.</au><au>Lo, Chung‐Mau</au><au>Poon, Ronnie T.P.</au><au>Fan, Sheung‐Tat</au><au>Man, Kwan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Suppression of tumorigenesis and metastasis of hepatocellular carcinoma by shRNA interference targeting on homeoprotein Six1</atitle><jtitle>International journal of cancer</jtitle><addtitle>Int J Cancer</addtitle><date>2010-08-15</date><risdate>2010</risdate><volume>127</volume><issue>4</issue><spage>859</spage><epage>872</epage><pages>859-872</pages><issn>0020-7136</issn><eissn>1097-0215</eissn><coden>IJCNAW</coden><abstract>We previously demonstrated that the overexpression of homeoprotein Six1 in hepatocellular carcinoma (HCC) patients is associated with venous infiltration, advanced pathologic tumor metastasis (pTNM) stage and poor overall survival rate (Ng et al. Br J Cancer 2006;95:1050–5). In this study, short hairpin RNA (shRNA) interference approach was used to suppress the expression of Six1 in a metastatic HCC cell line MHCC97L. Stable transfectant MHCC97L‐shSix1 carrying Six1‐specific shRNA plasmid was established to downregulate Six1 expression to about 40% when compared with MHCC97L‐Control. In vitro functional assays demonstrated that the growth rate and proliferation ability of MHCC97L‐shSix1 cells were markedly decreased. Moreover, significant decrease of cell motility and invasiveness were observed in MHCC97L‐shSix1 cells. Data from in vivo xenograft tumorigenesis model demonstrated that the size of tumor in MHCC97L‐shSix1 group was dramatically reduced. Experimental and spontaneous metastasis models indicated that targeting Six1 suppression noticeably reduced the pulmonary metastasis in MHCC97L‐shSix1 group. To identify Six1‐regulated targets, cDNA microarray was employed to compare the expression profiles of MHCC97L‐Control and MHCC97L‐shSix1 cells. Twenty‐eight downregulated and 24 upregulated genes with known functions were identified in MHCC97L‐shSix1. The functions of these target genes are involved in diverse biological activities. Our data suggest that Six1 may be involved in regulation of proliferation and invasiveness of HCC; thus targeting suppression of Six1 is a viable option for treating HCC patients.</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>20013809</pmid><doi>10.1002/ijc.25105</doi><tpages>14</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Biological and medical sciences Biomarkers, Tumor - genetics Biomarkers, Tumor - metabolism Blotting, Western Carcinoma, Hepatocellular - genetics Carcinoma, Hepatocellular - prevention & control Carcinoma, Hepatocellular - secondary cDNA microarray Cell Adhesion Cell Cycle Cell Movement Cell Proliferation Colony-Forming Units Assay Fluorescent Antibody Technique Gastroenterology. Liver. Pancreas. Abdomen Gene Expression Profiling Gene Expression Regulation, Neoplastic hepatocellular carcinoma Homeodomain Proteins - antagonists & inhibitors Homeodomain Proteins - genetics Homeodomain Proteins - metabolism homeoprotein Six1 Humans interference Kidney Neoplasms - genetics Kidney Neoplasms - prevention & control Kidney Neoplasms - secondary Liver Neoplasms, Experimental - genetics Liver Neoplasms, Experimental - pathology Liver Neoplasms, Experimental - prevention & control Liver. Biliary tract. Portal circulation. Exocrine pancreas Medical sciences metastasis Mice Mice, Nude Oligonucleotide Array Sequence Analysis Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger - genetics RNA, Messenger - metabolism RNA, Small Interfering - pharmacology short hairpin RNA (shRNA) Splenic Neoplasms - genetics Splenic Neoplasms - prevention & control Splenic Neoplasms - secondary Tumors Wound Healing |
| title | Suppression of tumorigenesis and metastasis of hepatocellular carcinoma by shRNA interference targeting on homeoprotein Six1 |
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