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中华放射肿瘤学杂志
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中华放射肿瘤学杂志  2021, Vol. 30 Issue (1): 90-94    DOI: 10.3760/cma.j.cn113030-20190510-00173
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沉默lncRNA HOTAIR上调miR-17-5p表达增加U87R细胞放射敏感性
原高明, 孟晓锋, 郭孝龙, 程小兵, 郝晓伟, 史保中
河南科技大学第一附属医院神经外科,洛阳 471000
Silencing lncRNA HOTAIR increases radiosensitivity of glioma cells by up-regulating miR-17-5p expression
Yuan Gaoming, Meng Xiaofeng, Guo Xiaolong, Cheng Xiaobing, Hao Xiaowei, Shi Baozhong
Department of Neurosurgery,First Affiliated Hospital of Henan University of Science and Technology,Luoyang 471000,China
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摘要 目的 探讨lncRNA HOTAIR对胶质瘤U87R细胞和移植瘤放射敏感性影响及潜在作用机制。方法 将阴性对照质粒、沉默HOTAIR质粒、过表达miR-NC质粒、过表达miR-17-5p质粒分别转染到U87R细胞中,记为沉默对照组、沉默HOTAIR组、过表达miR-NC组、过表达miR-17-5p组;以上各组细胞分别用4Gy照射,记为沉默对照+4Gy组、沉默HOTAIR+4Gy组、过表达miR-NC+4Gy组、过表达miR-17-5p+4Gy组;将沉默HOTAIR质粒分别与抑制表达miR-NC质粒、抑制表达miR-17-5p质粒共转染到U87R细胞中,记为沉默HOTAIR+抑制miR-NC组、沉默HOTAIR+抑制miR-17-5p组,转染均用脂质体法。采用qRT-PCR检测miR-17-5p和HOTAIR的表达;细胞克隆形成实验检测瘤细胞放射敏感性影响;流式细胞术检测细胞凋亡;双荧光素酶报告基因检测实验检测荧光活性。结果 HOTAIR在放射抵抗的细胞中高表达;沉默HOTAIR和过表达miR-17-5p可增加U87R细胞放射敏感性且促进放射照射诱导的凋亡。HOTAIR可靶向调节miR-17-5p表达,抑制miR-17-5p逆转了沉默HOTAIR对U87R细胞放射增敏和促进放射诱导的凋亡。结论 沉默lncRNA HOTAIR对胶质瘤细胞具有放射增敏和促放射诱导的凋亡作用,其机制可能与调控miR-17-5p有关。
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作者相关文章
原高明
孟晓锋
郭孝龙
程小兵
郝晓伟
史保中
关键词 HOX转录物反义基因miR-17-5p基因放射敏感性U87R细胞系    
AbstractObjective To investigate the effect of lncRNA HOTAIR on the radiosensitivity of glioma cells and its underlying mechanism. Methods The negative control plasmid,HOTAIR silencing plasmid,miR-NC over expressing plasmid,miR-17-5p over expressing plasmid were transfected into U87R cells,and assigned intothe silencing control,HOTAIR silencing,miR-NC over expressing and miR-17-5 pover expressing groups. Cells in the the above groups were irradiated at a dose of 4Gy,and recorded as silencing control+4Gy group,HOTAIRsilencing+4Gy group,miR-NC over expressing+4Gy group and miR-17-5p over expressing+4Gy group. The HOTAIR silencing plasmid,miR-NC suppressing plasmid and miR-17-5p suppressing plasmid were co-transfected into U87R cells and recorded as the HOTAIR silencing+miR-NCsuppressing group and HOTAIR silencing+miR-17-5p suppressing group. All procedures were transfected by the liposome method. The expression of miR-17-5p and HOTAIR was detected by qRT-PCR. The radio sensitivity of glioma cells was evaluated by cell clone formation assay. The cell apoptosis was assessed by flow cytometry. The fluorescence activity was assessed by dual luciferase reporter assay.Results HOTAIR was highly expressed in the radiation-resistant glioma cells. Silencing HOTAIR and over-expressing miR-17-5p could increase the radiosensitivity of U87R cells and promote radiation-induced apoptosis of U87R cells. HOTAIR could target and regulate the miR-17-5p expression. Suppressing miR-17-5p reversed the effect of silencing HOTAIR on U87R cell sensitization and promoting radiation-induced U87R cell apoptosis. Conclusions Silencing lncRNA HOTAIR yields radiation sensitization and promotes radiation-induced apoptosis in glioma cells. The mechanism may be related to the regulation of miR-17-5p.
Key wordsHOX transcript antisense gene    miR-17-5pgene    Radiosensitivity    U87R cell line   
收稿日期: 2019-05-10     
通讯作者: 史保中,Email:Luoyangxq@126.com   
引用本文:   
原高明,孟晓锋,郭孝龙等. 沉默lncRNA HOTAIR上调miR-17-5p表达增加U87R细胞放射敏感性[J]. 中华放射肿瘤学杂志, 2021, 30(1): 90-94.
Yuan Gaoming,Meng Xiaofeng,Guo Xiaolong et al. Silencing lncRNA HOTAIR increases radiosensitivity of glioma cells by up-regulating miR-17-5p expression[J]. Chinese Journal of Radiation Oncology, 2021, 30(1): 90-94.
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http://journal12.magtechjournal.com/Jweb_fszlx/CN/10.3760/cma.j.cn113030-20190510-00173     或     http://journal12.magtechjournal.com/Jweb_fszlx/CN/Y2021/V30/I1/90
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