文章摘要
李 科,刘明华,张 雷,徐世伟,尹昌林,任小宝,向 强.MicroRNA-21靶向调控β-catenin促进骨肉瘤细胞U2OS的增殖与侵袭[J].,2019,19(13):2448-2452
MicroRNA-21靶向调控β-catenin促进骨肉瘤细胞U2OS的增殖与侵袭
Role of miR-21 in the Proliferation and Invasion of Osteosarcoma U2OS Cells by Regulating β-catenin Expression
投稿时间:2018-12-28  修订日期:2019-01-23
DOI:10.13241/j.cnki.pmb.2019.13.010
中文关键词: 骨肉瘤  微小RNA-21  β-连环蛋白  增殖  侵袭
英文关键词: Osteosarcoma  Micro RNA-21  β-catenin  Proliferation  Invasion
基金项目:国家自然科学基金项目(81071537)
作者单位E-mail
李 科 解放军第三军医大学西南医院急诊科 重庆 400038 583702829@qq.com 
刘明华 解放军第三军医大学西南医院急诊科 重庆 400038  
张 雷 解放军第三军医大学西南医院急诊科 重庆 400038  
徐世伟 解放军第三军医大学西南医院急诊科 重庆 400038  
尹昌林 解放军第三军医大学西南医院急诊科 重庆 400038  
任小宝 解放军第三军医大学西南医院急诊科 重庆 400038  
向 强 解放军第三军医大学西南医院急诊科 重庆 400038  
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中文摘要:
      摘要 目的:研究微小RNA-21(microRNA-21,miR-21)对骨肉瘤细胞U2OS增殖与侵袭的影响及其可能机制。方法:采用Real-time PCR (RT-PCR)检测miR-21在骨肉瘤和临近正常骨组织中的表达差异。通过脂质体转染法将miR-21模拟物(microRNA-21 mimics,即mimics 组)及microRNA 无关序列(microRNA-NC,即NC组)转染入骨肉瘤细胞U2OS,real-time PCR(RT-PCR)检测miR-21和β-catenin mRNA 在U2OS细胞中的表达,Western blot检测β-catenin蛋白在U2OS细胞中的表达,并通过双荧光素酶报告基因验证miR-21与β-catenin基因3'-非编码区(3'-untranslated region,3'-UTR)的特异性结合作用。MTT法检测U2OS细胞体外增殖能力;Transwell侵袭模型探查U2OS细胞侵袭潜能。结果:骨肉瘤组织中miR-21水平显著高于正常骨组织(P<0.05)。过表达miR-21能够增强细胞增殖与侵袭,上调U2OS细胞β-catenin mRNA 和蛋白的表达。双荧光素酶报告基因结果表明miR-21可与β-catenin基因3'-UTR结合,从而对β-catenin的表达起调控作用。结论:miR-21可能通过调节β-catenin的表达促进骨肉瘤细胞U2OS的增殖与侵袭。
英文摘要:
      ABSTRACT Objective: To investigate the role of microRNA-21 (miR-21) in the proliferation and invasion potential of osteosarcoma cell lines U2OS and the exact underlying mechanism. Methods: The expression level of miR-21 in human osteosarcoma tissues and adjacent normal bone tissues was detected by real-time PCR (qRT-PCR). Osteosarcoma cells U2OS were transfected with chemically-synthesized miR-21 mimics and miR-NC, the expression of miR-21 and β-catenin were determined by qRT-PCR and western blot. The specific binding ability of miR-21 to β-catenin 3'-UTR was theoretically predicted and detected by the dual luciferase reporter gene assay. Moreover, the proliferation and invasion of U2OS cells were detected by MTT and Transwell assay. Results: The expression levels of miR-21 was higher in human osteosarcoma tissues compared to adjacent normal bone tissues (P<0.05). miR-21 directly up-regulates the mRNA and protein expression levels of β-catenin (P<0.05) and promotes proliferation and invasion abilities of U2OS cells. Luciferase reporter gene assay showed that miR-21 targeted the 3'-UTR of β-catenin and regulated the expression of β-catenin. Conclusion: miR-21 could enhance the proliferation and invasion abilities of osteosarcoma cell lines U2OS via upregulating the β-catenin expression.
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