人端粒酶启动子联合 miR-34a 整合靶向系统在胶质瘤细胞中的表达及对增殖的影响  

Targeted Efficacies of hTERT and miR-34a Integration System for Glioma

作　　者：张金灿[1] 胡丹 贺礼进[1] 桂成佳[1] 

机构地区：[1]湖南省永州市人民医院神经外科,湖南永州425100

出　　处：《医学临床研究》2015年第3期486-488,共3页Journal of Clinical Research

摘　　要：【目的】探讨人端粒酶启动子（hT ERT ）联合正常组织特异性miR‐34a （M IR）的整合靶向系统（hT ERT‐M IR）对胶质瘤细胞靶向性的影响。【方法】首先检测hT ERT、miR‐34a在人胶质瘤细胞株 U87、H4和人胶质细胞株HEB（正常细胞株）中的表达；通过质粒构建分别获得 hTERT‐Luc、hTERT‐MIR‐Luc、hTERT‐Bax 以及hTERT‐MIR‐Bax表达质粒，将hTERT‐Luc、hTERT‐MIR‐Luc质粒分别转染U87、H4和 HEB细胞中，检测三组细胞中hT ERT 与hT ERT‐M IR载体的表达活性；将hT ERT‐Bax以及hT ERT‐M IR‐Bax质粒分别转染U87、H4和HEB细胞中，比较三组细胞中hTERT‐Bax和hTERT‐MIR‐Bax对胶质瘤细胞增殖抑制能力的影响。【结果】hTERT在人胶质瘤细胞株U87和H4中特异性高表达，miR‐34a在人胶质细胞株HEB中特异性高表达；在正常胶质细胞株 HEB中，hTERT‐MIR的活性较 hTERT 明显减低（ P ＜0．01）；在胶质瘤细胞株 U87和 H4h中TERT‐Bax和hTERT‐MIR‐Bax 对细胞增殖抑制比较差异无显著性，但在正常胶质细胞株 HEB中，hTERT‐M IR‐Bax对细胞的增殖抑制明显小于hT ERT‐Bax。【结论】hT ERT‐M IR是胶质瘤基因靶向治疗的有效载体系统。[Objective]To explore a novel vector system based on hTERT promoter and cell‐specific miR‐34a for targeted therapy of glioma .[Methods] The expression levels of hTERT and miR‐34a were detected by quanti‐tative real‐time polymerase chain reaction （qRT‐PCR） in U87 and H4 glioma and normal colloid cells .The vectors of hTERT‐Luc ,hTERT‐MIR‐Luc ,hTERT‐Bax and hTERT‐MIR‐Bax were constructed and transfected into U 87 and H4 glioma and normal colloid cells respectively .The expression activities of hTERT and hTERT‐MIR vectors were detected by luciferase assay in U 87 and H4 glioma and normal colloid cells .And the proliferative capacities of U87 and H4 glioma and normal colloid cells regulated by hTERT‐Bax and hTERT‐MIR‐Bax were evaluated by thi‐azolyl blue tetrazolium blue （MTT） .[Results]The expression of hTERT promoter was stronger in U87 and H4 cells and miR‐34a was highly expressed in normal colloid cells .After incorporating normal cell‐specific miRNA‐targeted sequences ,the activity of hTERT‐MIR system decreased in normal cells （ P 〈0 .01） .Furthermore ,the proliferative capacity of glioma cells after transfection showed no significant differences .However ,significant differences existed in normal colloid cells .[Conclusion]hTERT‐MIR is an effective vector system of targeted gene therapy for glioma .

关 键 词：端粒  末端转移酶/遗传学 基因表达 启动区(遗传学) 微 RNAS 神经胶质瘤/遗传学 细胞增殖 

分 类 号：R338.15[医药卫生—人体生理学]