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下调miR-21可以增强食管癌TE-1细胞的放射敏感性 被引量：8

Down-regulation of miR-21 expression enhances the radiosensitivity of TE-1 cells in vitro
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摘要 目的研究miR-21下调对食管癌TE-1细胞的放射敏感性的影响。方法采用慢病毒介导的方法将含有反义miR21表达基因的载体转染至食管癌TE-1细胞系,嘌呤霉素筛选稳定下调miR21的食管癌细胞亚系,命名为TE-1-miR21^-;荧光定量PCR检测TE-1-miR21^-中miR21的相对表达量;细胞克隆形成实验测定TE-1和TE-1-miR21^-细胞的放射敏感性;RT-PCR和Western-blot法测定TE-1和TE-1-miR21^-细胞中β-catenin的变化;流式细胞术检测TE-1和TE-1-miR21^-细胞中p75NTR^+细胞的比例。结果成功构建稳定下调miR21的食管癌细胞亚系TE-1-miR21^-,荧光定量PCR结果显示miR21表达量明显下调;细胞克隆形成实验提示TE-1-miR21^-细胞较TE-1细胞的放射敏感性明显增强;RT-PCR结果分析显示,TE-1-miR21^-细胞与TE-1细胞的β-catenin mRNA的表达量无明显差异;Western blot的结果分析显示,TE-1-miR21^-细胞在接受高剂量照射(8Gy,10Gy)时,β-catenin蛋白量明显下降;流式细胞仪分析结果显示,TE-1-miR21^-细胞中p75NTR^+细胞的比例较TE-1细胞明显降低。结论下调miR21可以增强食管癌TE-1细胞的放射敏感性;下调-miR21增强放射敏感性的机制可能与wnt/β-catenin信号通路的活化程度降低以及食管癌TE-1细胞中p75NTR^+细胞比例减少有关。 Objective To study the effect of miR-21 down-regulation on the radiosensitivity of TE-1 ceils in vitro. Methods TE-1 cells were transfected via lentivirus with a vector containing the antisense oligonucleotides of miR21, and the subclones with stable down-regulation of miR21 expression were selected with puromycin and designated as TE-l-miR21-, whose expression level of miR21 was determined using real-time quantitative PCR. The radiosensitivity of TE-1 and TE-1-miR21^- cells were evaluated with colony formation assay, and the expressions of β-catenin was determined using Western blotting and RT-PCR. Flow cytometry was used to analyze the proportion of p75NTR^＋ cells in TE-1 and TE-1-miR21 cells. Results A cell subclone stably expressing a low level of miR21 was obtained and verified by real-time quantitative PCR. Colony formation assay showed an enhanced the radiosensitivity of TE-1-miR21^- cells compared to parental TE-1 ceils. RT-PCR revealed no significant changes in β-catenin mRNA expression in TE-1-miR21^- ceils, whereas its β-catenin protein expression was markedly suppressed by high-dose （8 and 10 Gy） irradiation. Flow cytometry assay showed a decreased proportion of p75NTR^＋ cells in TE-1-miR21 cells compared to that in TE-1 cells. Conclusion Down-regulation of miR21 can enhance the radiosensitivity of TE-1 cells, which might result from the inactivation of wnt/β-catenin signal pathway and a decreased p75NTR^＋ cell proportion.
作者 李晓青 陈鑫 黄珊 车少敏 张晓智
机构地区 西安交通大学第一附属医院放疗科
出处 《南方医科大学学报》 CAS CSCD 北大核心 2012年第11期1559-1563,共5页 Journal of Southern Medical University
基金 国家自然科学基金(30972962)~~
关键词 食管癌 放射敏感性 MIR-21 WNT Β-CATENIN信号通路 P75NTR esophageal cancer radiosensitivity miR-21 wnt/β-catenin pathway p75NTR
分类号 R735.1 [医药卫生—肿瘤]
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参考文献16

  • 1殷蔚伯,谷铣之肿瘤放射治疗学[M].3版.北京:北京医科大学中国协和医科大学联合出版社,2008:598-620.
  • 2Che SM, Zhang XZ, Hou L, et al. Cyclooxygenase-2 inhibitor NS398 enhances radiosensitivity of radioresistant esophageal cancer cells by inhibiting AKT activation and inducing apoptosis [J]. Cancer Invest, 2010, 28(7): 679-88.
  • 3Tiscornia G, Singer O, Verma IM. Production and purification of lentiviral vectors[J]. Nat Protoc, 2006, 1 (1): 241-5.
  • 4Yanaihara N, Caplen N, Bowman E, et al. Unique microRNA molecular profilesin lung cancer diagnosis and prognosis [J]. Cancer Cell, 2006, 9(3): 189-98.
  • 5Volinia S, Calin GA, Liu CG, et al. A microRNA expression signature of human solid tumors defines cancer gene targets [J]. Acad, 2006, 103(7): 2257-61.
  • 6Feber A, Xi L, Luketich JD, et al. MicroRNA expression profiles of esophageal cancer [J]. Thorac Cardiovasc Surg, 2008, 135(2): 255-60.
  • 7Tran N, McLean T, Zhang X, et al. MicroRNA expression profiles in head and neck cancer cell lines [J]. Biochem. Biophys Res Commun, 2007, 358(1): 12-7.
  • 8Sempere FL, Christensen M, Silahtaroglu A, et al. Altered micro- RNA expression confined to specific epithelial cell subpopulations in breast cancer[J]. Cancer Res, 2007, 67(24): 11612-20.
  • 9Mori Y, Ishiguro H, Kuwabara Y, et al. MicroRNA-21 induces cell proliferation and invasion in esophageal squamous cell carcinoma [J]. Mol Med Report, 2009, 2(2): 235-9.
  • 10Akagi I, Miyashita M, Ishibashi O, et al. Relationship between altered expression levels of MIR21, MIR143, MIR145, and MIR205 and elinicopathologic features of esophageal squamous cell carcinoma[J]. Dis Esophagus, 2011, 24(7): 523-30.

同被引文献89

  • 1Bartel DP. MicroRNAs: target recognition and regulatory functions [J]. Ce11,2009,136(2):215-233.
  • 2Lee YS, Dutta A. MicroRNAs in cancer [ J ]. Annu Rev Pathol, 2009,4 : 199 - 227.
  • 3Iorio MV, Croce CM. MicroRNA dysregulation in cancer:diagnos- tics, monitoring and therapeutics: A comprehensive review [ J ]. EMBo Mol Med,2012,4 (3) : 143 - 159.
  • 4David PB. MicroRNAs: genonfies, biogenesis, mechanism, and function [ J ]. Ce11,2004,116 ( 2 ) : 281 - 297.
  • 5Chen S,Wang H,NQ WL,et al. Radiosensitizing effects of ectopic miR - 101 on Non - Small - Cell Lung cancer cells depend on the endogenous miR - 101 level [ J]. Radiation Oncology Biol Phys, 2011,81 (5) :1524 - 1529.
  • 6Qu C, Liang ZH, Huang JL, et al. MiR - 205 determines the ra- dioresistance of human nasopharyngeal carcinoma by directly targe- ting PTEN [ J ]. Cell Cycle,2012,11 (4) :785 - 796.
  • 7Lin J ,Liu C ,Gao F, et al. miR -200c Enhances radiosensitivity of human breast cancer cell [ J ]. J Cell Biochem, 2013,114 : 606 - 615.
  • 8Calin GA, Sevignani C, Dumitru CD, et al. Human microRNA Genes are frequently located at fragile sites and genomic regions in- volved in cancers[J]. Proc Natl Acad Sci U S A,2004,101(9) : 2999 - 3004.
  • 9Chan JK, Blansit K, Kiet T, et al. The inhibition of miR-21 pro- motes apoptosis and chemosensitivity in ovarian cancer[ J]. Gynecol Oncol 2014, pii: S0090 -8258( 14)00094 -8.
  • 10Petrovic N, Mandusic V, Stanojevic B, et al. The difference in miR - 21 expression levels between invasive and non - invasive breast cancer emphasizes its role in breast cancer invasion [ J ]. Med Oncol,2014,31 ( 3 ) :867.

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南方医科大学学报
2012年 第11期

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