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我们发现,一种在RNA干扰实验中用作阴性对照的商业化siRNA具有明显的诱导人慢性髓性白血病K562细胞系向红系方向分化的作用.它表现为K562细胞瞬时转染该siRNA后,红系分化的特异表面标志CD235及ε-、γ-和β-珠蛋白的表达升高,GATA-2的表达降低,细胞增殖速度减慢,软琼脂克隆形成率降低,并且此过程不伴随细胞凋亡.而生物信息学分析显示,该siRNA序列与目前所有已知人类基因均无明显同源性.研究结果提示,该siRNA不适于用作红系分化实验中的阴性对照.siRNA的作用远比人们目前所知的要复杂得多,siRNA的脱靶效应应当引起研究者的足够重视,在RNA干扰实验中阴性对照siRNA的选择会极大地影响对实验结果的判读.
We found that a commercial siRNA used as a negative control in RNA interference experiments has a distinct effect on inducing erythroid differentiation of human chronic myelogenous leukemia K562 cell line.It appears that after transient transfection of this siRNA into K562 cells, Erythroid differentiation specific surface markers CD235 and ε-, γ- and β-globin expression increased, GATA-2 expression decreased, the rate of cell proliferation slowed, soft agar colony formation rate decreased, and the process is not accompanied by cells Apoptosis.Analysis of bioinformatics showed that the siRNA sequence had no significant homology with all the known human genes at present.These results suggest that the siRNA is not suitable as a negative control in erythroid differentiation experiments.The role of siRNA is far It is much more complicated than people know now. The off-target effect of siRNA should attract enough attention from researchers. The choice of negative control siRNA in RNA interference experiments will greatly affect the interpretation of experimental results.