{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2024,6,10]],"date-time":"2024-06-10T19:40:16Z","timestamp":1718048416158},"reference-count":23,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2015,8,26]],"date-time":"2015-08-26T00:00:00Z","timestamp":1440547200000},"content-version":"vor","delay-in-days":25,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"Natural science foundation of Fujin Province","award":["2010D017"],"award-info":[{"award-number":["2010D017"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Orthopaedic Surgery"],"published-print":{"date-parts":[[2015,8]]},"abstract":"<jats:sec><jats:title>Objective<\/jats:title><jats:p>To investigate the influence of knocking down ezrin expression in combination with heat shock protein (<jats:styled-content style=\"fixed-case\">HSP<\/jats:styled-content>)\u2010induced immune killing on the apoptosis and proliferation of mouse osteosarcoma cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The HSP70 and ezrin\u2010shRNA DNA fragments cloned into the expression vector pGFP\u2010V\u2010RS and the expression vectors pGFP\u2010V\u2010RS\u2010shRNA and pGFP\u2010V\u2010RS\u2010shRNA\u2010HSP70 constructed and transfected into MG63 cell line, where their status was observed by fluorescent microscopy. Expression of ezrin and HSP70 was determined by RT\u2010PCR and western blot. Changes in cell apoptosis and proliferation were assessed by flow cytometry and MTS and changes in expression of apoptosis and cell cycle\u2010related proteins by western blot. Specific cytotoxic T lymphocytes (CTLs) were induced by HSP70 and its lethal effect on target MG63 tumor cells analyzed by MTS assay.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The specific vector simultaneously downregulated ezrin and upregulated<jats:styled-content style=\"fixed-case\">HSP<\/jats:styled-content>70. Compared with ezrin knockdown alone, simultaneous<jats:styled-content style=\"fixed-case\">HSP<\/jats:styled-content>70 overexpression partially recovered the promoted cellular apoptosis and proliferation suppression by induced by ezrin knockdown; however, the apoptosis rate of<jats:styled-content style=\"fixed-case\">MG<\/jats:styled-content>63 cells was significantly greater than that of a negative control. In addition, ezrin\u2010<jats:styled-content style=\"fixed-case\">shRNA<\/jats:styled-content>and ezrin\u2010<jats:styled-content style=\"fixed-case\">shRNA<\/jats:styled-content>\/<jats:styled-content style=\"fixed-case\">HSP<\/jats:styled-content>70 promoted expression of<jats:styled-content style=\"fixed-case\">B<\/jats:styled-content>ax. However, expression of these agents reduces<jats:styled-content style=\"fixed-case\">B<\/jats:styled-content>cl\u20102 and<jats:styled-content style=\"fixed-case\">C<\/jats:styled-content>yclin<jats:styled-content style=\"fixed-case\">D<\/jats:styled-content>1.<jats:styled-content style=\"fixed-case\">T<\/jats:styled-content>he cytotoxic effects of<jats:styled-content style=\"fixed-case\">CTLs<\/jats:styled-content>on target<jats:styled-content style=\"fixed-case\">MG<\/jats:styled-content>63 tumor cells were significantly greater in the<jats:styled-content style=\"fixed-case\">CTL<\/jats:styled-content>+<jats:styled-content style=\"fixed-case\">IL<\/jats:styled-content>\u20102 +<jats:styled-content style=\"fixed-case\">HSP<\/jats:styled-content>70 group than the<jats:styled-content style=\"fixed-case\">CTL<\/jats:styled-content>+<jats:styled-content style=\"fixed-case\">IL<\/jats:styled-content>\u20102 group.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Simultaneously knocking down ezrin and overexpressing<jats:styled-content style=\"fixed-case\">HSP<\/jats:styled-content>70 promotes apoptosis and inhibits proliferation of osteosarcoma cells and<jats:styled-content style=\"fixed-case\">HSP<\/jats:styled-content>70 induces<jats:styled-content style=\"fixed-case\">CTL,<\/jats:styled-content>enhancing the lethal effect on tumor cells.<\/jats:p><\/jats:sec>","DOI":"10.1111\/os.12186","type":"journal-article","created":{"date-parts":[[2015,8,27]],"date-time":"2015-08-27T03:59:40Z","timestamp":1440647980000},"page":"273-280","source":"Crossref","is-referenced-by-count":5,"title":["Effects of Ezrin and Heat Shock Protein 70 on Apoptosis and Proliferation of Human Osteosarcoma Cells"],"prefix":"10.1111","volume":"7","author":[{"given":"Qin","family":"Yao","sequence":"first","affiliation":[{"name":"Central Laboratory Xiamen China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hui\u2010yi","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Spine Surgery Zhongshan Hospital Xiamen University Xiamen China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bo\u2010zhen","family":"Xie","sequence":"additional","affiliation":[{"name":"Department of Spine Surgery Zhongshan Hospital Xiamen University Xiamen China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2015,8,26]]},"reference":[{"key":"e_1_2_5_2_1","first-page":"1027","article-title":"Preliminary report of a randomized controlled multicenter study of first\u2010line chemotherapy regimen combined with angiogenesis inhibitor for osteosarcoma of the extremities","volume":"32","author":"Shi X","year":"2012","journal-title":"Zhonghua Gu Ke Za 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