摘要
目的 研究RNA干扰(RNA interference, RNAi) 对白血病细胞株WT1 基因表达的抑制作用.探讨WT1小干扰RNA在白血病治疗中的应用.方法 设计制备多对针对WT1基因的小干扰RNA(small interfering RNA, siRNA),转染HL-60人白血病细胞株,采用RT-PCR法测定WT1 mRNA 的表达,Western blot 测定WT1 蛋白的表达,流式细胞仪检测细胞凋亡率.结果 siRNA 后WT1 mRNA与蛋白的表达强度分别为(23.62±1.28)%和(23.20±1.04)%,和对照组比较有显著性差异(P＜0.05);在WT1表达受到抑制的同时,HL-60细胞的凋亡率为(13.72 ±1.33)%,和对照组比较有显著性差异(P＜0.05).结论 siRNA能有效抑制HL-60细胞中WT1基因的表达,增加细胞凋亡.
Abstract
Objective To investigate the inhibitory effect of RNA interference (RNAi) on WT1 gene expression in HL-60 cell line. Methods Several pairs of small interfering RNA ( siRNA) for WT1 gene was designed and transfected into HL-60 cells. WT1 gene expression was assessed by RT-PCR, and WT1 protein expression was tested by Western blot. Flow cytometry was used to measure the cell apoptosis. Results The expression rates of WT1 mRNA and the expression rates of WT1 protein were (23.62% ± 1. 28% ) , (23.20% ± 1.04% ) , respectively in HL-60 WT1 siRNA-transfected cells, significantly lower than those of control cells (P <0. 05) ; The apoptosis rates were (13. 72% ± 1. 33% ) , significantly higher than those of control cells (P <0. 05). Conclusion Expression of WT1 gene can be effectively inhibited by siRNA in HL-60 cells. The siRNA targeting human WT1 is considered to be possible tool against leukemia.
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