miR-21海绵吸附载体在脑胶质瘤细胞中的应用  

The application of miRNAs-21 sponge adsorption vector in brain glioma cells

作　　者：陈波[1] 陈丹祎[1] 徐文中[2] 

机构地区：[1]陕西省商洛市中心医院神经外科,陕西商洛726000 [2]河南科技大学大学第二附属神经外科,河南洛阳471000

出　　处：《中华神经外科疾病研究杂志》2014年第1期9-12,共4页Chinese Journal of Neurosurgical Disease Research

摘　　要：目的探讨微小RNA-21(miR-21)海绵吸附载体在脑胶质瘤细胞中发挥的作用。方法采用microRNA(miRNA)海绵技术构建人源miR-21(has-miR-21)反义片段(miR-21AS),并串联重复8个构建至真核表达载体(pCMV),转染脑胶质瘤细胞U251,实时荧光定量聚合酶链式反应(qRTPCR)检测miR-21的表达,四甲基偶氨唑盐(MTT)实验检测细胞增殖能力,并检测miR-21肿瘤密切相关靶基因肿瘤抑制基因原肌球蛋白1(tumor suppressor gene tropomyosin 1,TPM1)、第10号染色体缺失的磷酸酶和张力蛋白同源基因(phosphate and tension homology deleted on chromsome ten,PTEN)、生长素β受体(transforming growth factor-beta,TGF-β)、人MutS蛋白同基因2(Human mutS homolog2,hMSH2)、程序性细胞死亡4基因(programmed cell death protein 4,PDCD4)的表达。结果 U251细胞转染重组pCMV-miR-21AS质粒后显著下调了miRNA-21的水平,也显著降低了细胞增殖能力,显著提高了TPM1、PTEN、TGF-β、hMSH2、PDCD4的mRNA水平。结论构建的miR-21海绵吸附载体可显著下调miR-21水平,从而降低肿瘤细胞学效应,为肿瘤miRNA基因治疗提供理论基础。Objective To investigate the role of miRNA-21 （nfiR-21） sponge adsorption vector in brain gliorm cells. Methods microRNA （miRNA） sponge technology was used to build has-miR-21 antisense fi＇agment and transfect the brain glioma cell line U251. miR-21 expression was detected by real rime polymerase chain reactions （qRT-PCR）, cell proliferation capacity was detected by dimethyl thiazolyl diphenyl tetrazolium （MTI＇）, and the expressions of tumor suppressor tropomyosin 1 （TPM1）, phosphate and tension homology deleted on chromsome ten （PTEN）, transforming growth factor-beta （TGF-13）, Human mutS homolog 2 （hMSH2）, programmed cell death protein 4 （PI^X）4） were detected. Results pCMV-miR-21AS tmmfeeted U251 cells significantly down-regulated the level of miR-21 and lowered the cell proliferation, and increased the mRNA levels of TPM1, PTEN, TGF-β, hMSI-12, PDCD4. Conclusion miR-21 sponge adsorption vector can down-regulate the level of miR-21, thereby reducing the tumor cytological effects, which paves the way for miRNA gene therapy in the tuinor.

关 键 词：微小RNA-21 海绵吸附载体 脑胶质瘤细胞 

分 类 号：R743[医药卫生—神经病学与精神病学]