RNAi技术沉默K562细胞中c-myc基因的初步研究

c-myc gene silencing in K562 cells with RNA interference

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。

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.