miR-218-5p inhibits the stem cell properties and invasive ability of the A2B5+CD133− subgroup of human glioma stem cells
MicroRNAs (miRs) act as oncogenes or tumor-suppressor genes, and regulate the proliferation, apoptosis, invasion, differentiation, angiogenesis and behavior of glioma stem cells, which are important in glioma development and recurrence. The present study was performed to investigate the impact of mi...
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| Veröffentlicht in: | Oncology reports 2016-02, Vol.35 (2), p.869-877 |
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| creator | WU, ZHIWU HAN, YONG LI, YANYAN LI, XUETAO SUN, TING CHEN, GUILIN HUANG, YULUN ZHOU, YOUXIN DU, ZIWEI |
| description | MicroRNAs (miRs) act as oncogenes or tumor-suppressor genes, and regulate the proliferation, apoptosis, invasion, differentiation, angiogenesis and behavior of glioma stem cells, which are important in glioma development and recurrence. The present study was performed to investigate the impact of miR-218-5p on stem cell properties and invasive ability of the A2B5+CD133− human glioma stem cell subgroup. qRT-PCR was used to detect miR-218-5p expression in non-cancerous brain and human glioma tissues, human glioma cell lines and human glioma stem cell lines. Lentivirus vectors encoding miR-218-5p and anti-miR-218-5p were constructed and stably transfected into A2B5+CD133− SHG-139s cells. Neurosphere formation Cell Counting Kit-8 (CCK-8) and Transwell assays, immunofluorescence and qRT-PCR analyses were used to explore the role of miR-218-5p in SHG-139s cells. qRT-PCR analysis showed that miR-218-5p expression was lower in human glioma tissues and cells than in non-cancerous brain tissues and normal human astrocyte cells, and lower in A2B5+CD133− (SHG-139s) cells than in CD133+ (SU2 and U87s) cells. The CCK-8 assay demonstrated that the growth curve was significantly inhibited in the miR-218-5p-SHG-139s cells compared to the miR-control, blank and anti-miR-218-5p groups. The neurosphere formation assay indicated that upregulation of miR-218-5p expression inhibited SHG-139s neurosphere formation. Immunofluorescence staining and qRT-PCR showed that miR-218-5p reduced stem cell marker (A2B5, nestin, PLAGL2, ALDH1 and Sox2) expression compared with the controls; however, immunofluorescence staining analysis showed that upregulation of miR-218-5p expression led to no difference in CD133 expression. miR-218-5p reduced SHG-139s cell invasiveness in the Transwell assay and reduced MMP9 expression as detected in qRT-PCR and immunofluorescence analyses. All differences were statistically significant. miR-218-5p expression was lower in human glioma tissues, cells and the A2B5+CD133− human glioma stem cell subgroup. miR-218-5p may be a tumor-suppressor gene in glioma that functions by upregulating miR-218-5p expression, which inhibits the stem cell properties and invasive properties of SHG-139s cells. |
| doi_str_mv | 10.3892/or.2015.4418 |
| format | Article |
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The present study was performed to investigate the impact of miR-218-5p on stem cell properties and invasive ability of the A2B5+CD133− human glioma stem cell subgroup. qRT-PCR was used to detect miR-218-5p expression in non-cancerous brain and human glioma tissues, human glioma cell lines and human glioma stem cell lines. Lentivirus vectors encoding miR-218-5p and anti-miR-218-5p were constructed and stably transfected into A2B5+CD133− SHG-139s cells. Neurosphere formation Cell Counting Kit-8 (CCK-8) and Transwell assays, immunofluorescence and qRT-PCR analyses were used to explore the role of miR-218-5p in SHG-139s cells. qRT-PCR analysis showed that miR-218-5p expression was lower in human glioma tissues and cells than in non-cancerous brain tissues and normal human astrocyte cells, and lower in A2B5+CD133− (SHG-139s) cells than in CD133+ (SU2 and U87s) cells. The CCK-8 assay demonstrated that the growth curve was significantly inhibited in the miR-218-5p-SHG-139s cells compared to the miR-control, blank and anti-miR-218-5p groups. The neurosphere formation assay indicated that upregulation of miR-218-5p expression inhibited SHG-139s neurosphere formation. Immunofluorescence staining and qRT-PCR showed that miR-218-5p reduced stem cell marker (A2B5, nestin, PLAGL2, ALDH1 and Sox2) expression compared with the controls; however, immunofluorescence staining analysis showed that upregulation of miR-218-5p expression led to no difference in CD133 expression. miR-218-5p reduced SHG-139s cell invasiveness in the Transwell assay and reduced MMP9 expression as detected in qRT-PCR and immunofluorescence analyses. All differences were statistically significant. miR-218-5p expression was lower in human glioma tissues, cells and the A2B5+CD133− human glioma stem cell subgroup. miR-218-5p may be a tumor-suppressor gene in glioma that functions by upregulating miR-218-5p expression, which inhibits the stem cell properties and invasive properties of SHG-139s cells.</description><identifier>ISSN: 1021-335X</identifier><identifier>EISSN: 1791-2431</identifier><identifier>DOI: 10.3892/or.2015.4418</identifier><identifier>PMID: 26572167</identifier><language>eng</language><publisher>Greece: D.A. Spandidos</publisher><subject>A2B5+CD133− glioma stem cells ; Adult ; Brain Neoplasms - genetics ; Brain Neoplasms - pathology ; Brain research ; Cancer therapies ; Cell Line, Tumor ; Cell Proliferation - genetics ; Chemotherapy ; Cloning ; Female ; Fluorescent Antibody Technique ; Gene Expression Regulation, Neoplastic - genetics ; Gene Knockdown Techniques ; Genes ; Genes, Tumor Suppressor ; Genotype & phenotype ; Glioma ; Glioma - genetics ; Glioma - pathology ; Growth factors ; Humans ; invasive ability ; Laboratories ; Male ; Medical research ; MicroRNAs - genetics ; Middle Aged ; miR-218-5p ; Neoplasm Invasiveness ; Neoplastic Stem Cells - pathology ; Patients ; Penicillin ; Polymerase Chain Reaction ; stem cell properties ; Stem cells ; Studies ; Surgery ; Transfection ; Tumors</subject><ispartof>Oncology reports, 2016-02, Vol.35 (2), p.869-877</ispartof><rights>Copyright: © Wu et al.</rights><rights>Copyright Spandidos Publications UK Ltd. 2016</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c318t-4d8f5a22d9a45f10ef4b7b153a2c790b42ee470ce892303e86bfdfb56cc210b93</citedby><cites>FETCH-LOGICAL-c318t-4d8f5a22d9a45f10ef4b7b153a2c790b42ee470ce892303e86bfdfb56cc210b93</cites></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/26572167$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>WU, ZHIWU</creatorcontrib><creatorcontrib>HAN, YONG</creatorcontrib><creatorcontrib>LI, YANYAN</creatorcontrib><creatorcontrib>LI, XUETAO</creatorcontrib><creatorcontrib>SUN, TING</creatorcontrib><creatorcontrib>CHEN, GUILIN</creatorcontrib><creatorcontrib>HUANG, YULUN</creatorcontrib><creatorcontrib>ZHOU, YOUXIN</creatorcontrib><creatorcontrib>DU, ZIWEI</creatorcontrib><title>miR-218-5p inhibits the stem cell properties and invasive ability of the A2B5+CD133− subgroup of human glioma stem cells</title><title>Oncology reports</title><addtitle>Oncol Rep</addtitle><description>MicroRNAs (miRs) act as oncogenes or tumor-suppressor genes, and regulate the proliferation, apoptosis, invasion, differentiation, angiogenesis and behavior of glioma stem cells, which are important in glioma development and recurrence. The present study was performed to investigate the impact of miR-218-5p on stem cell properties and invasive ability of the A2B5+CD133− human glioma stem cell subgroup. qRT-PCR was used to detect miR-218-5p expression in non-cancerous brain and human glioma tissues, human glioma cell lines and human glioma stem cell lines. Lentivirus vectors encoding miR-218-5p and anti-miR-218-5p were constructed and stably transfected into A2B5+CD133− SHG-139s cells. Neurosphere formation Cell Counting Kit-8 (CCK-8) and Transwell assays, immunofluorescence and qRT-PCR analyses were used to explore the role of miR-218-5p in SHG-139s cells. qRT-PCR analysis showed that miR-218-5p expression was lower in human glioma tissues and cells than in non-cancerous brain tissues and normal human astrocyte cells, and lower in A2B5+CD133− (SHG-139s) cells than in CD133+ (SU2 and U87s) cells. The CCK-8 assay demonstrated that the growth curve was significantly inhibited in the miR-218-5p-SHG-139s cells compared to the miR-control, blank and anti-miR-218-5p groups. The neurosphere formation assay indicated that upregulation of miR-218-5p expression inhibited SHG-139s neurosphere formation. Immunofluorescence staining and qRT-PCR showed that miR-218-5p reduced stem cell marker (A2B5, nestin, PLAGL2, ALDH1 and Sox2) expression compared with the controls; however, immunofluorescence staining analysis showed that upregulation of miR-218-5p expression led to no difference in CD133 expression. miR-218-5p reduced SHG-139s cell invasiveness in the Transwell assay and reduced MMP9 expression as detected in qRT-PCR and immunofluorescence analyses. All differences were statistically significant. miR-218-5p expression was lower in human glioma tissues, cells and the A2B5+CD133− human glioma stem cell subgroup. miR-218-5p may be a tumor-suppressor gene in glioma that functions by upregulating miR-218-5p expression, which inhibits the stem cell properties and invasive properties of SHG-139s cells.</description><subject>A2B5+CD133− glioma stem cells</subject><subject>Adult</subject><subject>Brain Neoplasms - genetics</subject><subject>Brain Neoplasms - pathology</subject><subject>Brain research</subject><subject>Cancer therapies</subject><subject>Cell Line, Tumor</subject><subject>Cell Proliferation - genetics</subject><subject>Chemotherapy</subject><subject>Cloning</subject><subject>Female</subject><subject>Fluorescent Antibody Technique</subject><subject>Gene Expression Regulation, Neoplastic - genetics</subject><subject>Gene Knockdown Techniques</subject><subject>Genes</subject><subject>Genes, Tumor Suppressor</subject><subject>Genotype & phenotype</subject><subject>Glioma</subject><subject>Glioma - genetics</subject><subject>Glioma - pathology</subject><subject>Growth factors</subject><subject>Humans</subject><subject>invasive ability</subject><subject>Laboratories</subject><subject>Male</subject><subject>Medical research</subject><subject>MicroRNAs - genetics</subject><subject>Middle Aged</subject><subject>miR-218-5p</subject><subject>Neoplasm Invasiveness</subject><subject>Neoplastic Stem Cells - pathology</subject><subject>Patients</subject><subject>Penicillin</subject><subject>Polymerase Chain Reaction</subject><subject>stem cell properties</subject><subject>Stem cells</subject><subject>Studies</subject><subject>Surgery</subject><subject>Transfection</subject><subject>Tumors</subject><issn>1021-335X</issn><issn>1791-2431</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNpd0d9r1TAUB_AgipvTN58lIIjgcs05SdrmcV5_TBgIouBbSdp0N6NtuqQdzL_A5_2J_iVLvXOCTwnkk8M550vIc-AbUWl8G-IGOaiNlFA9IIdQamAoBTzMd47AhFA_DsiTlC44x5IX-jE5wEKVCEV5SH4O_itDqJiaqB933vo50XnnaJrdQBvX93SKYXJx9i5RM7ZZXZnkrxw11vd-vqah-_PhBN-pN9v3IMTvXzc0LfY8hmVaX3fLYEZ63vswmH9101PyqDN9cs_uziPy_eOHb9tTdvbl0-ftyRlrBFQzk23VKYPYaiNVB9x10pYWlDDYlJpbic7JkjcuL0Nw4arCdm1nVdE0CNxqcURe7-vmQS4Xl-Z68GntwIwuLKmGUqFQgFxm-vI_ehGWOObuatACi0pqpbI63qsmhpSi6-op-sHE6xp4vWZSh1ivmdRrJpm_uCu62MG19_hvCBm82oM05QX7NqR7EyITinFkvCq0uAUrJZMF</recordid><startdate>20160201</startdate><enddate>20160201</enddate><creator>WU, ZHIWU</creator><creator>HAN, YONG</creator><creator>LI, YANYAN</creator><creator>LI, XUETAO</creator><creator>SUN, TING</creator><creator>CHEN, GUILIN</creator><creator>HUANG, YULUN</creator><creator>ZHOU, YOUXIN</creator><creator>DU, ZIWEI</creator><general>D.A. 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genetics</topic><topic>Brain Neoplasms - pathology</topic><topic>Brain research</topic><topic>Cancer therapies</topic><topic>Cell Line, Tumor</topic><topic>Cell Proliferation - genetics</topic><topic>Chemotherapy</topic><topic>Cloning</topic><topic>Female</topic><topic>Fluorescent Antibody Technique</topic><topic>Gene Expression Regulation, Neoplastic - genetics</topic><topic>Gene Knockdown Techniques</topic><topic>Genes</topic><topic>Genes, Tumor Suppressor</topic><topic>Genotype & phenotype</topic><topic>Glioma</topic><topic>Glioma - genetics</topic><topic>Glioma - pathology</topic><topic>Growth factors</topic><topic>Humans</topic><topic>invasive ability</topic><topic>Laboratories</topic><topic>Male</topic><topic>Medical research</topic><topic>MicroRNAs - genetics</topic><topic>Middle Aged</topic><topic>miR-218-5p</topic><topic>Neoplasm Invasiveness</topic><topic>Neoplastic Stem Cells - pathology</topic><topic>Patients</topic><topic>Penicillin</topic><topic>Polymerase Chain Reaction</topic><topic>stem cell properties</topic><topic>Stem cells</topic><topic>Studies</topic><topic>Surgery</topic><topic>Transfection</topic><topic>Tumors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>WU, ZHIWU</creatorcontrib><creatorcontrib>HAN, YONG</creatorcontrib><creatorcontrib>LI, YANYAN</creatorcontrib><creatorcontrib>LI, XUETAO</creatorcontrib><creatorcontrib>SUN, TING</creatorcontrib><creatorcontrib>CHEN, GUILIN</creatorcontrib><creatorcontrib>HUANG, YULUN</creatorcontrib><creatorcontrib>ZHOU, YOUXIN</creatorcontrib><creatorcontrib>DU, ZIWEI</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>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</collection><collection>ProQuest One Community College</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>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>Oncology reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>WU, ZHIWU</au><au>HAN, YONG</au><au>LI, YANYAN</au><au>LI, XUETAO</au><au>SUN, TING</au><au>CHEN, GUILIN</au><au>HUANG, YULUN</au><au>ZHOU, YOUXIN</au><au>DU, ZIWEI</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>miR-218-5p inhibits the stem cell properties and invasive ability of the A2B5+CD133− subgroup of human glioma stem cells</atitle><jtitle>Oncology reports</jtitle><addtitle>Oncol Rep</addtitle><date>2016-02-01</date><risdate>2016</risdate><volume>35</volume><issue>2</issue><spage>869</spage><epage>877</epage><pages>869-877</pages><issn>1021-335X</issn><eissn>1791-2431</eissn><abstract>MicroRNAs (miRs) act as oncogenes or tumor-suppressor genes, and regulate the proliferation, apoptosis, invasion, differentiation, angiogenesis and behavior of glioma stem cells, which are important in glioma development and recurrence. The present study was performed to investigate the impact of miR-218-5p on stem cell properties and invasive ability of the A2B5+CD133− human glioma stem cell subgroup. qRT-PCR was used to detect miR-218-5p expression in non-cancerous brain and human glioma tissues, human glioma cell lines and human glioma stem cell lines. Lentivirus vectors encoding miR-218-5p and anti-miR-218-5p were constructed and stably transfected into A2B5+CD133− SHG-139s cells. Neurosphere formation Cell Counting Kit-8 (CCK-8) and Transwell assays, immunofluorescence and qRT-PCR analyses were used to explore the role of miR-218-5p in SHG-139s cells. qRT-PCR analysis showed that miR-218-5p expression was lower in human glioma tissues and cells than in non-cancerous brain tissues and normal human astrocyte cells, and lower in A2B5+CD133− (SHG-139s) cells than in CD133+ (SU2 and U87s) cells. The CCK-8 assay demonstrated that the growth curve was significantly inhibited in the miR-218-5p-SHG-139s cells compared to the miR-control, blank and anti-miR-218-5p groups. The neurosphere formation assay indicated that upregulation of miR-218-5p expression inhibited SHG-139s neurosphere formation. Immunofluorescence staining and qRT-PCR showed that miR-218-5p reduced stem cell marker (A2B5, nestin, PLAGL2, ALDH1 and Sox2) expression compared with the controls; however, immunofluorescence staining analysis showed that upregulation of miR-218-5p expression led to no difference in CD133 expression. miR-218-5p reduced SHG-139s cell invasiveness in the Transwell assay and reduced MMP9 expression as detected in qRT-PCR and immunofluorescence analyses. All differences were statistically significant. miR-218-5p expression was lower in human glioma tissues, cells and the A2B5+CD133− human glioma stem cell subgroup. miR-218-5p may be a tumor-suppressor gene in glioma that functions by upregulating miR-218-5p expression, which inhibits the stem cell properties and invasive properties of SHG-139s cells.</abstract><cop>Greece</cop><pub>D.A. Spandidos</pub><pmid>26572167</pmid><doi>10.3892/or.2015.4418</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | A2B5+CD133− glioma stem cells Adult Brain Neoplasms - genetics Brain Neoplasms - pathology Brain research Cancer therapies Cell Line, Tumor Cell Proliferation - genetics Chemotherapy Cloning Female Fluorescent Antibody Technique Gene Expression Regulation, Neoplastic - genetics Gene Knockdown Techniques Genes Genes, Tumor Suppressor Genotype & phenotype Glioma Glioma - genetics Glioma - pathology Growth factors Humans invasive ability Laboratories Male Medical research MicroRNAs - genetics Middle Aged miR-218-5p Neoplasm Invasiveness Neoplastic Stem Cells - pathology Patients Penicillin Polymerase Chain Reaction stem cell properties Stem cells Studies Surgery Transfection Tumors |
| title | miR-218-5p inhibits the stem cell properties and invasive ability of the A2B5+CD133− subgroup of human glioma stem cells |
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