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目的:探讨不同白血病细胞株及不同亚型急性白血病患者原代骨髓细胞中WT1(17AA+)剪接变异体的表达水平。方法:建立实时定量RT-PCR方法,检测K562、SHI、Jurkat、NB4、NB4／WTIA、NB4／WTID等白血病细胞株及79例初诊急性白血病患者原代骨髓细胞中WT1基因和WT1(17AA+)剪接变异体表达水平,并计算WT1(17AA+)／WT1比值。结果:白血病细胞株K562、SH1、NB4、NB4／WT1A、NB4／WT1D中WT1基因表达水平较Jurkat、U937高2～3个对数级,且K562和NB4细胞株中WT1表达以WT1(17AA+)剪接变异体优势表达。79例急性白血病患者原代骨髓细胞中WT1总体表达水平在0．03-34．17之间,中位数为1．02,明显高于23例对照非白血病患者(P<0．001),其WT1(17AA+)／WT1比值在0．30～0．93之间,中位数为0．62,WT1(17AA+)剪接变异体表达在不同亚型急性白血病之间无统计学差异(F=0．152, P=0．979)。结论:高表达WT1的白血病细胞株及白血病患者骨髓细胞中WT1基因以WT1(17AA+)剪接变异体优势表达,且WT1(17AA+)剪接变异体表达在不同亚型急性白血病之间无明显统计学差别。
Objective: To investigate the expression of WT1 (17AA +) splicing variant in primary leukemic cells of different leukemia cell lines and different subtypes of acute leukemia. Methods: Real-time quantitative RT-PCR was used to detect WT1 (17AA +) splicing in primary myeloid cells from K562, SHI, Jurkat, NB4, NB4 / WTIA and NB4 / WTID leukemia cell lines and 79 newly diagnosed acute leukemia patients Variant expression levels and WT1 (17AA +) / WT1 ratio were calculated. Results WT1 gene expression in leukemia cell lines K562, SH1, NB4, NB4 / WT1A and NB4 / WT1D was 2-3 logarithmically higher than that of Jurkat and U937, and WT1 (17AA +) was expressed in K562 and NB4 cell lines Splice variants predominantly expressed. The expression level of WT1 in primary bone marrow cells of 79 patients with acute leukemia was between 0.03-34.17, with a median of 1.02, which was significantly higher than that of 23 non-leukemia patients (P <0.001) The WT1 (17AA +) / WT1 ratio was between 0.30 and 0.93 with a median of 0.62. There was no significant difference in WT1 (17AA +) splice variant expression between different subtypes of acute leukemia (F = 0.152, P = 0.979). CONCLUSION: WT1 gene is highly expressed in WT1 (17AA +) splicing variant in leukemia cells and leukemia patients with high expression of WT1, and WT1 (17AA +) splicing variant has no significant difference in different subtypes of acute leukemia .