转染靶向hTERT miRNA表达框架对K562细胞端粒酶活性及增殖活性的影响  

Discussing the inhibition effect of mi RNA expression cassettes targeting h TERT on telomerase activity and K562 proliferation activity

作　　者：赵丹丹[1] 徐慧[1] 李亚红[1] 凌晗 吴珊珊[1] 羊金菊 彭剑雄[1] 

机构地区：[1]中南大学湘雅医学院医学检验系,湖南长沙410013

出　　处：《南京医科大学学报（自然科学版）》2016年第3期293-297,306,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：湖南省大学生研究性学习和创新性实验计划项目(CY14277);国家自由探索项目(201510533374);国家研究性学习和创新性实验计划项目(201510533026)

摘　　要：目的 :设计靶向端粒酶逆转录酶(telomerase reverse transcriptase,h TERT)的mi RNA表达框架,探讨其对h TERT的表达、端粒酶活性及K562增殖的影响,以期为慢性髓系白血病提供新的基因治疗策略。方法:构建靶向h TERT的mi RNA表达框架,脂质体法转染K562细胞,银染法及荧光定量PCR法检测端粒酶活性及h TERT的表达情况。Annexin V/PI双染检测细胞凋亡。结果:mi RNA表达框架转染48 h后,h TERT的表达明显下降,端粒酶活性明显减低,K562细胞凋亡率增加。结论:靶向h TERT的mi RNA表达框架能够有效干扰h TERT的表达,抑制端粒酶活性,诱导K562细胞的凋亡。以mi RNA表达框架为基础的RNAi技术,有望成为分子水平治疗慢性髓系白血病的有效工具。Objective：To design the mi RNA expression cassettes targeting telomerase reverse transcriptase（h TERT）and to discuss the interfering effect on the expression of h TERT,the telomerase activity and the proliferation of K562,so as to provide a new gene therapy for chronic myeloid leukemia. Methods：mi RNA expression cassettes targeting h TERT were constructed and transfected to K562 cells by liposome transfection. The telomerase activity and the expression of h TERT were detected by TRAP-silver staining and q PCR,respectively. Cell apoptosis rate was detected by Annexin V / PI. Results：After 48 h transfection,the expression of h TERT and telomerase activity were significantly reduced. The apoptosis rate of K562 cell was increased. Conclusion：The mi RNA expression cassettes targeting h TERT can effectively interfere with the expression of h TERT,inhibit the telomerase activity and induce the apoptosis of K562 cells. RNAi technology based on the mi RNA expression cassette is expected to become an efficient tool in molecular level for the treatment of chronic leukemia.

关 键 词：MIRNA RNAi 端粒酶 HTERT 慢性髓系白血病 

分 类 号：R730.5[医药卫生—肿瘤]