南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (06): 647-650.

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RNAi技术沉默K562细胞中c-myc基因的初步研究

岳枫1, 马文丽1, 宋艳斌1, 石嵘1, 张宝1, 郑文岭2   

  1. 1. 南方医科大学分子生物学研究所, 广东, 广州, 510515;
    2. 广州军区广州总医院医学实验科, 广东, 广州, 510010
  • 出版日期:2005-06-20 发布日期:2005-06-20
  • 基金资助:
    收稿日期:2004-10-26。
    基金项目:国家自然科学基金(39880032)
    作者简介:岳枫(1975- ),女,在读硕士研究生,电话:020-61640114-89097,E-mail:yuefeng@fimmu.com
    通讯作者:马文丽,电话:020-61648210,E-mail:Wenli@fimmu.com

c-myc gene silencing in K562 cells with RNA interference

YUE Feng1, MA Wen-li1, SONG Yan-bin1, SHI Rong1, ZHANG Bao1, ZHENG Wen-ling2   

  1. 1. 南方医科大学分子生物学研究所, 广东, 广州, 510515;
    2. 广州军区广州总医院医学实验科, 广东, 广州, 510010
  • Online:2005-06-20 Published:2005-06-20

摘要: 目的 运用RNAi技术,设计针对c-myc的小干扰RNA(siRNA),研究其干扰效果。方法 针对c-mycmRNA的第1357靶位点设计siRNAs,经LipofectamineTM2000脂质体法转染K562细胞,进行RT-PCR、细胞计数和MTT法及流式细胞仪检测,观察其干扰效果。结果 转染组与阴性对照组和空白对照组相比,c-mycmRNA表达明显减弱,细胞计数和MTTD(λ)值均有显著降低,并有明显的凋亡率。结论 运用RNAi技术,可以有效地干扰c-myc的表达,并进一步诱导细胞凋亡。

Abstract: Objective To study the inhibitory effect of small interfering RNA (siRNA) targeting c-myc gene in K562 cells. Methods siRNAs targeting the site 1357 of c-myc mRNA was designed and synthesized. In vitro cultured K562 cells were transfected with lipofectamineTM 2000 and the inhibitory effect was detected by reverse transcriptase (RT)-PCR, cell count, MTT assay and fluorescence-activated cell sorting. Results Compared with the negative and blank control group, the transfection group showed marked decrease in the c-myc expression and the K562 cells exhibited increased apoptosis rate. Conclusion RNA interference can effectively inhibit c-myc expression and induce apoptosis in K562 cells.

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