白血病细胞系WT1的表达及反义核苷酸对其的抑制作用

目的:探讨RT-PCR方法测定白血病细胞系WT1基因的表达以及WT1反义寡核苷酸对白血病细胞系增殖的抑制作用。方法:应用RT-PCR方法测定K562、HL-60、TF-1及U937 4株白血病细胞系WT1基因的表达,然后应用针对WT1翻译起始位点的反义寡核苷酸(WT1 ASO)处理K562细胞系、U937细胞系,采用锥虫蓝拒染法确定白血病细胞的增殖。结果:K562、HL-60及TF-1 3株白血病细胞系均高度表达WT1,而U937细胞系未测到WT1。WT1 ASO能够抑制WT1表达阳性的K562细胞的生长,对WT1表达阴性的U937则无抑制作用;而WT1有义寡核苷酸对K562细胞系、U937细胞系均无抑制作用。结论:RT-PCR测定方法可以检测WT1基因在白血病细胞系表达,WT1反义寡核苷酸对白血病细胞有特异的抑制作用。WT1基因在白血病细胞增殖过程中起一定作用。

Expression of Wilms' tumor gene (WT1) in leukemia lines and the inhibitory effect of WT1 antisense oligonucleotide on leukemia lines

Objective:To elucidate the expression of Wilm's tumor gene (WT1) in leukemia lines and the inhibitory effect of WTl antisense oligonucleotide on proliferation of leukemia cells lines. Method: Expression of WT1 mRNA was detected in four leukemia cell lines by using RT-PCR. K562, U937 cell lines were treated with WT1 antisense oligonucleotide(WT1 ASO) targeted to the translation site of WTl mRNA, then the inhibitory effect on growth of leukemia cells lines was measured by Trypan Blue exclusion method. Result: High expression of WTl mRNA was found in K562,HL-60 and TF-1 cell lines, but not in U937 cell lines. WT1 ASO significantly inhibited the proliferation of K562 cell line not expressing WT1, but didn't inhibit the growth of U937 cells. Conclusion: WT1 gene is expressed in majority of leukemia cell lines, WTl ASO has the specific inhibitory effect on growth of leukemic cells lines expressing WT1, and WT1 may play an important role in proliferation of leukemic cells.