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MicroRNA-221 Induces Cell Survival and Cisplatin Resistance through PI3K/Akt Pathway in Human Osteosarcoma

BACKGROUND: MicroRNAs are short regulatory RNAs that negatively modulate protein expression at a post-transcriptional and/or translational level and are deeply involved in the pathogenesis of several types of cancers. Specifically, microRNA-221 (miR-221) is overexpressed in many human cancers, where...

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Autores principales: Zhao, Guangyi, Cai, Chengkui, Yang, Tongtao, Qiu, Xiuchun, Liao, Bo, Li, Wei, Ji, Zhenwei, Zhao, Jian, Zhao, Haien, Guo, Mingjun, Ma, Qiong, Xiao, Chun, Fan, Qingyu, Ma, Baoan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3553141/
https://www.ncbi.nlm.nih.gov/pubmed/23372675
http://dx.doi.org/10.1371/journal.pone.0053906
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author Zhao, Guangyi
Cai, Chengkui
Yang, Tongtao
Qiu, Xiuchun
Liao, Bo
Li, Wei
Ji, Zhenwei
Zhao, Jian
Zhao, Haien
Guo, Mingjun
Ma, Qiong
Xiao, Chun
Fan, Qingyu
Ma, Baoan
author_facet Zhao, Guangyi
Cai, Chengkui
Yang, Tongtao
Qiu, Xiuchun
Liao, Bo
Li, Wei
Ji, Zhenwei
Zhao, Jian
Zhao, Haien
Guo, Mingjun
Ma, Qiong
Xiao, Chun
Fan, Qingyu
Ma, Baoan
author_sort Zhao, Guangyi
collection PubMed
description BACKGROUND: MicroRNAs are short regulatory RNAs that negatively modulate protein expression at a post-transcriptional and/or translational level and are deeply involved in the pathogenesis of several types of cancers. Specifically, microRNA-221 (miR-221) is overexpressed in many human cancers, wherein accumulating evidence indicates that it functions as an oncogene. However, the function of miR-221 in human osteosarcoma has not been totally elucidated. In the present study, the effects of miR-221 on osteosarcoma and the possible mechanism by which miR-221 affected the survival, apoptosis, and cisplatin resistance of osteosarcoma were investigated. METHODOLOGY/PRINCIPAL FINDINGS: Real-time quantitative PCR analysis revealed miR-221 was significantly upregulated in osteosarcoma cell lines than in osteoblasts. Both human osteosarcoma cell lines SOSP-9607 and MG63 were transfected with miR-221 mimic or inhibitor to regulate miR-221 expression. The effects of miR-221 were then assessed by cell viability, cell cycle analysis, apoptosis assay, and cisplatin resistance assay. In both cells, upregulation of miR-221 induced cell survival and cisplatin resistance and reduced cell apoptosis. In addition, knockdown of miR-221 inhibited cell growth and cisplatin resistance and induced cell apoptosis. Potential target genes of miR-221 were predicted using bioinformatics. Moreover, luciferase reporter assay and western blot confirmed that PTEN was a direct target of miR-221. Furthermore, introduction of PTEN cDNA lacking 3′-UTR or PI3K inhibitor LY294002 abrogated miR-221-induced cisplatin resistance. Finally, both miR-221 and PTEN expression levels in osteosarcoma samples were examined by using real-time quantitative PCR and immunohistochemistry. High miR-221 expression level and inverse correlation between miR-221 and PTEN levels were revealed in osteosarcoma tissues. CONCLUSIONS/SIGNIFICANCE: These results for the first time demonstrate that upregulation of miR-221 induces the malignant phenotype of human osteosarcoma whereas knockdown of miR-221 reverses this phenotype, suggesting that miR-221 could be a potential target for osteosarcoma treatment.
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spelling pubmed-35531412013-01-31 MicroRNA-221 Induces Cell Survival and Cisplatin Resistance through PI3K/Akt Pathway in Human Osteosarcoma Zhao, Guangyi Cai, Chengkui Yang, Tongtao Qiu, Xiuchun Liao, Bo Li, Wei Ji, Zhenwei Zhao, Jian Zhao, Haien Guo, Mingjun Ma, Qiong Xiao, Chun Fan, Qingyu Ma, Baoan PLoS One Research Article BACKGROUND: MicroRNAs are short regulatory RNAs that negatively modulate protein expression at a post-transcriptional and/or translational level and are deeply involved in the pathogenesis of several types of cancers. Specifically, microRNA-221 (miR-221) is overexpressed in many human cancers, wherein accumulating evidence indicates that it functions as an oncogene. However, the function of miR-221 in human osteosarcoma has not been totally elucidated. In the present study, the effects of miR-221 on osteosarcoma and the possible mechanism by which miR-221 affected the survival, apoptosis, and cisplatin resistance of osteosarcoma were investigated. METHODOLOGY/PRINCIPAL FINDINGS: Real-time quantitative PCR analysis revealed miR-221 was significantly upregulated in osteosarcoma cell lines than in osteoblasts. Both human osteosarcoma cell lines SOSP-9607 and MG63 were transfected with miR-221 mimic or inhibitor to regulate miR-221 expression. The effects of miR-221 were then assessed by cell viability, cell cycle analysis, apoptosis assay, and cisplatin resistance assay. In both cells, upregulation of miR-221 induced cell survival and cisplatin resistance and reduced cell apoptosis. In addition, knockdown of miR-221 inhibited cell growth and cisplatin resistance and induced cell apoptosis. Potential target genes of miR-221 were predicted using bioinformatics. Moreover, luciferase reporter assay and western blot confirmed that PTEN was a direct target of miR-221. Furthermore, introduction of PTEN cDNA lacking 3′-UTR or PI3K inhibitor LY294002 abrogated miR-221-induced cisplatin resistance. Finally, both miR-221 and PTEN expression levels in osteosarcoma samples were examined by using real-time quantitative PCR and immunohistochemistry. High miR-221 expression level and inverse correlation between miR-221 and PTEN levels were revealed in osteosarcoma tissues. CONCLUSIONS/SIGNIFICANCE: These results for the first time demonstrate that upregulation of miR-221 induces the malignant phenotype of human osteosarcoma whereas knockdown of miR-221 reverses this phenotype, suggesting that miR-221 could be a potential target for osteosarcoma treatment. Public Library of Science 2013-01-23 /pmc/articles/PMC3553141/ /pubmed/23372675 http://dx.doi.org/10.1371/journal.pone.0053906 Text en © 2013 Zhao et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Zhao, Guangyi
Cai, Chengkui
Yang, Tongtao
Qiu, Xiuchun
Liao, Bo
Li, Wei
Ji, Zhenwei
Zhao, Jian
Zhao, Haien
Guo, Mingjun
Ma, Qiong
Xiao, Chun
Fan, Qingyu
Ma, Baoan
MicroRNA-221 Induces Cell Survival and Cisplatin Resistance through PI3K/Akt Pathway in Human Osteosarcoma
title MicroRNA-221 Induces Cell Survival and Cisplatin Resistance through PI3K/Akt Pathway in Human Osteosarcoma
title_full MicroRNA-221 Induces Cell Survival and Cisplatin Resistance through PI3K/Akt Pathway in Human Osteosarcoma
title_fullStr MicroRNA-221 Induces Cell Survival and Cisplatin Resistance through PI3K/Akt Pathway in Human Osteosarcoma
title_full_unstemmed MicroRNA-221 Induces Cell Survival and Cisplatin Resistance through PI3K/Akt Pathway in Human Osteosarcoma
title_short MicroRNA-221 Induces Cell Survival and Cisplatin Resistance through PI3K/Akt Pathway in Human Osteosarcoma
title_sort microrna-221 induces cell survival and cisplatin resistance through pi3k/akt pathway in human osteosarcoma
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3553141/
https://www.ncbi.nlm.nih.gov/pubmed/23372675
http://dx.doi.org/10.1371/journal.pone.0053906
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