microRNA-524-5p inhibits proliferation and induces cell cycle arrest of osteosarcoma cells via targeting CDK6

Osteosarcoma (OS) is one of the most commonly diagnosed malignant tumors that mainly affects children and adolescents. The underlying molecular mechanisms that are responsible for the initiation and development of OS are still not clear. Increasing evidence suggested the tumor suppressor role of mic...

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Veröffentlicht in:Biochemical and biophysical research communications 2020-09, Vol.530 (3), p.566-573
Hauptverfasser: Chen, Hanwen, Cheng, Cai, Gao, Shuming
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Cheng, Cai
Gao, Shuming
description Osteosarcoma (OS) is one of the most commonly diagnosed malignant tumors that mainly affects children and adolescents. The underlying molecular mechanisms that are responsible for the initiation and development of OS are still not clear. Increasing evidence suggested the tumor suppressor role of microRNA-524-5p in a variety of cancers via targeting key pathways involved in tumorigenesis. The aim of this study was to characterize the function of miR-524-5p in OS. A total 50 paired OS tissues and adjacent normal tissues were collected from OS patients. The expression of miR-524-5p in OS tissues and cells was detected by RT-qPCR. The CCK-8 assay, flow cytometry and transwell assay were applied to determine the proliferation and invasion abilities of OS cells. The targets of miR-524-5p were predicted using the miRDB dataset and confirmed by luciferase reporter assay and western blot analysis. The expression of miR-524-5p was decreased in OS tissues and cell lines. OS patients with lymph node metastasis harbored relative lower level of miR-524-5p. Overexpression of miR-524-5p in OS cells significantly suppressed the proliferation, drove cell cycle arrest and apoptosis. The mechanism investigation revealed that miR-524-5p bound the 3′-untranslated region (UTR) of Cyclin Dependent Kinase 6 (CDK6) and repressed the expression of CDK6 in OS cells. Overexpressed CDK6 was found in OS tissues, which was inversely correlated with that of miR-524-5p. Moreover, forced expression of CDK6 significantly reversed the anti-cancer effects of miR-524-5p on the proliferation, apoptosis and cell cycle arrest of OS cells. Our results identified the tumor-suppressive role of miR-524-5p in OS via targeting CDK6, which may lead to the identification of novel therapeutic target for the treatment of OS. •miR-524-5p expression was decreased in OS tissues and cells.•Overexpressed miR-524-5p inhibited the proliferation, induced apoptosis and cell cycle arrest of OS cells.•miR-524-5p targeted CDK6 and induced cell cycle arrest in OS.•Restoration of CDK6 impaired the anti-tumor effects of miR-524-5p in OS.
doi_str_mv 10.1016/j.bbrc.2020.07.092
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The underlying molecular mechanisms that are responsible for the initiation and development of OS are still not clear. Increasing evidence suggested the tumor suppressor role of microRNA-524-5p in a variety of cancers via targeting key pathways involved in tumorigenesis. The aim of this study was to characterize the function of miR-524-5p in OS. A total 50 paired OS tissues and adjacent normal tissues were collected from OS patients. The expression of miR-524-5p in OS tissues and cells was detected by RT-qPCR. The CCK-8 assay, flow cytometry and transwell assay were applied to determine the proliferation and invasion abilities of OS cells. The targets of miR-524-5p were predicted using the miRDB dataset and confirmed by luciferase reporter assay and western blot analysis. The expression of miR-524-5p was decreased in OS tissues and cell lines. OS patients with lymph node metastasis harbored relative lower level of miR-524-5p. Overexpression of miR-524-5p in OS cells significantly suppressed the proliferation, drove cell cycle arrest and apoptosis. The mechanism investigation revealed that miR-524-5p bound the 3′-untranslated region (UTR) of Cyclin Dependent Kinase 6 (CDK6) and repressed the expression of CDK6 in OS cells. Overexpressed CDK6 was found in OS tissues, which was inversely correlated with that of miR-524-5p. Moreover, forced expression of CDK6 significantly reversed the anti-cancer effects of miR-524-5p on the proliferation, apoptosis and cell cycle arrest of OS cells. Our results identified the tumor-suppressive role of miR-524-5p in OS via targeting CDK6, which may lead to the identification of novel therapeutic target for the treatment of OS. •miR-524-5p expression was decreased in OS tissues and cells.•Overexpressed miR-524-5p inhibited the proliferation, induced apoptosis and cell cycle arrest of OS cells.•miR-524-5p targeted CDK6 and induced cell cycle arrest in OS.•Restoration of CDK6 impaired the anti-tumor effects of miR-524-5p in OS.</description><identifier>ISSN: 0006-291X</identifier><identifier>EISSN: 1090-2104</identifier><identifier>DOI: 10.1016/j.bbrc.2020.07.092</identifier><identifier>PMID: 32747087</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Adolescent ; Bone Neoplasms - genetics ; Bone Neoplasms - pathology ; CDK6 ; Cell Cycle Checkpoints ; Cell cycle progression ; Cell Proliferation ; Child ; Cyclin-Dependent Kinase 6 - genetics ; Gene Expression Regulation, Neoplastic ; Humans ; MicroRNAs - genetics ; miR-524-5p ; Osteosarcoma ; Osteosarcoma - genetics ; Osteosarcoma - pathology ; Tumor Cells, Cultured</subject><ispartof>Biochemical and biophysical research communications, 2020-09, Vol.530 (3), p.566-573</ispartof><rights>2020 Elsevier Inc.</rights><rights>Copyright © 2020 Elsevier Inc. 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The underlying molecular mechanisms that are responsible for the initiation and development of OS are still not clear. Increasing evidence suggested the tumor suppressor role of microRNA-524-5p in a variety of cancers via targeting key pathways involved in tumorigenesis. The aim of this study was to characterize the function of miR-524-5p in OS. A total 50 paired OS tissues and adjacent normal tissues were collected from OS patients. The expression of miR-524-5p in OS tissues and cells was detected by RT-qPCR. The CCK-8 assay, flow cytometry and transwell assay were applied to determine the proliferation and invasion abilities of OS cells. The targets of miR-524-5p were predicted using the miRDB dataset and confirmed by luciferase reporter assay and western blot analysis. The expression of miR-524-5p was decreased in OS tissues and cell lines. OS patients with lymph node metastasis harbored relative lower level of miR-524-5p. Overexpression of miR-524-5p in OS cells significantly suppressed the proliferation, drove cell cycle arrest and apoptosis. The mechanism investigation revealed that miR-524-5p bound the 3′-untranslated region (UTR) of Cyclin Dependent Kinase 6 (CDK6) and repressed the expression of CDK6 in OS cells. Overexpressed CDK6 was found in OS tissues, which was inversely correlated with that of miR-524-5p. Moreover, forced expression of CDK6 significantly reversed the anti-cancer effects of miR-524-5p on the proliferation, apoptosis and cell cycle arrest of OS cells. Our results identified the tumor-suppressive role of miR-524-5p in OS via targeting CDK6, which may lead to the identification of novel therapeutic target for the treatment of OS. •miR-524-5p expression was decreased in OS tissues and cells.•Overexpressed miR-524-5p inhibited the proliferation, induced apoptosis and cell cycle arrest of OS cells.•miR-524-5p targeted CDK6 and induced cell cycle arrest in OS.•Restoration of CDK6 impaired the anti-tumor effects of miR-524-5p in OS.</description><subject>Adolescent</subject><subject>Bone Neoplasms - genetics</subject><subject>Bone Neoplasms - pathology</subject><subject>CDK6</subject><subject>Cell Cycle Checkpoints</subject><subject>Cell cycle progression</subject><subject>Cell Proliferation</subject><subject>Child</subject><subject>Cyclin-Dependent Kinase 6 - genetics</subject><subject>Gene Expression Regulation, Neoplastic</subject><subject>Humans</subject><subject>MicroRNAs - genetics</subject><subject>miR-524-5p</subject><subject>Osteosarcoma</subject><subject>Osteosarcoma - genetics</subject><subject>Osteosarcoma - pathology</subject><subject>Tumor Cells, Cultured</subject><issn>0006-291X</issn><issn>1090-2104</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE1PGzEQhq2qqATKH-ih8rGXXcYfsTdSLygtHyICqSoSN8vrHaeOdtepvUHi39chlCOnOczzvpp5CPnCoGbA1Pmmbtvkag4catA1LPgHMmOwgIozkB_JDABUxRfs8Zic5LwBYEyqxSdyLLiWGho9I8MQXIq_7i6qOZfVfEvD-Ce0Ycp0m2IfPCY7hThSO3Zl1e0cZuqw76l7dj1SmxLmiUZPY54wZptcHOwLkelTsHSyaY1TGNd0-eNWfSZH3vYZz17nKXm4_Pl7eV2t7q9ulheryom5mirmpW40eqadE4CSe85cCw1gWSjRImtY10rVeSuFBgGOaS8aZxtlpW6tOCXfDr3lib-7cqEZQt4fZUeMu2y4FCBUw3VTUH5Ai4acE3qzTWGw6dkwMHvNZmP2ms1eswFtiuYS-vrav2sH7N4i_70W4PsBwPLlU8Bksgs4OuxCQjeZLob3-v8BF8-OUQ</recordid><startdate>20200924</startdate><enddate>20200924</enddate><creator>Chen, Hanwen</creator><creator>Cheng, Cai</creator><creator>Gao, Shuming</creator><general>Elsevier Inc</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>7X8</scope><orcidid>https://orcid.org/0000-0001-6586-1205</orcidid></search><sort><creationdate>20200924</creationdate><title>microRNA-524-5p inhibits proliferation and induces cell cycle arrest of osteosarcoma cells via targeting CDK6</title><author>Chen, Hanwen ; Cheng, Cai ; Gao, Shuming</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c356t-1f4787ef17cc30e42f21cb080ef4763be181db46dfa437030c17f38ca86a47ba3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Adolescent</topic><topic>Bone Neoplasms - genetics</topic><topic>Bone Neoplasms - pathology</topic><topic>CDK6</topic><topic>Cell Cycle Checkpoints</topic><topic>Cell cycle progression</topic><topic>Cell Proliferation</topic><topic>Child</topic><topic>Cyclin-Dependent Kinase 6 - genetics</topic><topic>Gene Expression Regulation, Neoplastic</topic><topic>Humans</topic><topic>MicroRNAs - genetics</topic><topic>miR-524-5p</topic><topic>Osteosarcoma</topic><topic>Osteosarcoma - genetics</topic><topic>Osteosarcoma - pathology</topic><topic>Tumor Cells, Cultured</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chen, Hanwen</creatorcontrib><creatorcontrib>Cheng, Cai</creatorcontrib><creatorcontrib>Gao, Shuming</creatorcontrib><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>Biochemical and biophysical research communications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chen, Hanwen</au><au>Cheng, Cai</au><au>Gao, Shuming</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>microRNA-524-5p inhibits proliferation and induces cell cycle arrest of osteosarcoma cells via targeting CDK6</atitle><jtitle>Biochemical and biophysical research communications</jtitle><addtitle>Biochem Biophys Res Commun</addtitle><date>2020-09-24</date><risdate>2020</risdate><volume>530</volume><issue>3</issue><spage>566</spage><epage>573</epage><pages>566-573</pages><issn>0006-291X</issn><eissn>1090-2104</eissn><abstract>Osteosarcoma (OS) is one of the most commonly diagnosed malignant tumors that mainly affects children and adolescents. The underlying molecular mechanisms that are responsible for the initiation and development of OS are still not clear. Increasing evidence suggested the tumor suppressor role of microRNA-524-5p in a variety of cancers via targeting key pathways involved in tumorigenesis. The aim of this study was to characterize the function of miR-524-5p in OS. A total 50 paired OS tissues and adjacent normal tissues were collected from OS patients. The expression of miR-524-5p in OS tissues and cells was detected by RT-qPCR. The CCK-8 assay, flow cytometry and transwell assay were applied to determine the proliferation and invasion abilities of OS cells. The targets of miR-524-5p were predicted using the miRDB dataset and confirmed by luciferase reporter assay and western blot analysis. The expression of miR-524-5p was decreased in OS tissues and cell lines. OS patients with lymph node metastasis harbored relative lower level of miR-524-5p. Overexpression of miR-524-5p in OS cells significantly suppressed the proliferation, drove cell cycle arrest and apoptosis. The mechanism investigation revealed that miR-524-5p bound the 3′-untranslated region (UTR) of Cyclin Dependent Kinase 6 (CDK6) and repressed the expression of CDK6 in OS cells. Overexpressed CDK6 was found in OS tissues, which was inversely correlated with that of miR-524-5p. Moreover, forced expression of CDK6 significantly reversed the anti-cancer effects of miR-524-5p on the proliferation, apoptosis and cell cycle arrest of OS cells. Our results identified the tumor-suppressive role of miR-524-5p in OS via targeting CDK6, which may lead to the identification of novel therapeutic target for the treatment of OS. •miR-524-5p expression was decreased in OS tissues and cells.•Overexpressed miR-524-5p inhibited the proliferation, induced apoptosis and cell cycle arrest of OS cells.•miR-524-5p targeted CDK6 and induced cell cycle arrest in OS.•Restoration of CDK6 impaired the anti-tumor effects of miR-524-5p in OS.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>32747087</pmid><doi>10.1016/j.bbrc.2020.07.092</doi><tpages>8</tpages><orcidid>https://orcid.org/0000-0001-6586-1205</orcidid></addata></record>
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subjects Adolescent
Bone Neoplasms - genetics
Bone Neoplasms - pathology
CDK6
Cell Cycle Checkpoints
Cell cycle progression
Cell Proliferation
Child
Cyclin-Dependent Kinase 6 - genetics
Gene Expression Regulation, Neoplastic
Humans
MicroRNAs - genetics
miR-524-5p
Osteosarcoma
Osteosarcoma - genetics
Osteosarcoma - pathology
Tumor Cells, Cultured
title microRNA-524-5p inhibits proliferation and induces cell cycle arrest of osteosarcoma cells via targeting CDK6
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