文章摘要
赵丹丹,徐 慧,李亚红,凌 晗,吴珊珊,羊金菊,彭剑雄.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[J].南京医科大学学报,2016,(3):293~297
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 miRNA expression cassettes targeting hTERT on telomerase activity and K562 proliferation activity
投稿时间:2015-11-18  
DOI:10.7655/NYDXBNS20160308
中文关键词: miRNA  RNAi  端粒酶  hTERT  慢性髓系白血病
英文关键词: miRNA  RNAi  telomerase  hTERT  chronic myeloid leukemia
基金项目: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)
作者单位
赵丹丹 中南大学湘雅医学院医学检验系,湖南 长沙 410013 
徐 慧 中南大学湘雅医学院医学检验系,湖南 长沙 410013 
李亚红 中南大学湘雅医学院医学检验系,湖南 长沙 410013 
凌 晗 中南大学湘雅医学院医学检验系,湖南 长沙 410013 
吴珊珊 中南大学湘雅医学院医学检验系,湖南 长沙 410013 
羊金菊 中南大学湘雅医学院医学检验系,湖南 长沙 410013 
彭剑雄 中南大学湘雅医学院医学检验系,湖南 长沙 410013 
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中文摘要:
      目的:设计靶向端粒酶逆转录酶(telomerase reverse transcriptase,hTERT)的miRNA表达框架,探讨其对hTERT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hTERT的miRNA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hTERT的表达情况。Annexin V/PI双染检测细胞凋亡。结果:miRNA表达框架转染48 h后,hTERT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hTERT的miRNA表达框架能够有效干扰hTERT的表达,抑制端粒酶活性,诱导K562细胞的凋亡。以miRNA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。
英文摘要:
      Objective:To design the miRNA expression cassettes targeting telomerase reverse transcriptase (hTERT)and to discuss the interfering effect on the expression of hTERT,the telomerase activity and the proliferation of K562,so as to provide a new gene therapy for chronic myeloid leukemia. Methods:miRNA expression cassettes targeting hTERT were constructed and transfected to K562 cells by liposome transfection. The telomerase activity and the expression of hTERT were detected by TRAP-silver staining and qPCR,respectively. Cell apoptosis rate was detected by Annexin V/PI. Results:After 48 h transfection,the expression of hTERT and telomerase activity were significantly reduced. The apoptosis rate of K562 cell was increased. Conclusion:The miRNA expression cassettes targeting hTERT can effectively interfere with the expression of hTERT,inhibit the telomerase activity and induce the apoptosis of K562 cells. RNAi technology based on the miRNA expression cassette is expected to become an efficient tool in molecular level for the treatment of chronic leukemia.
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