【摘要】 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。更多还原

【Abstract】 Objective:To investigate the expression pattern of Wilms tumor gene WT1(17AA+) isoform in different leukemia cell lines and primary bone marrow cells from patients with different subtypes of acute leukemia. Methods:The leukemia cell lines included K562, SHI, Jurkat, NB4, NB4/WT1A, and NB4/WT1D cells, etc. The primary bone marrow cells were collected from 79 patients initially diagnosed as acute leukemia. The expression levels of WT1 gene and WT1(17AA+) isoform were determined by real-time quantitative reverse transcriptase polymerase chain reaction (RQ-RT-PCR). The ratio of WT1(17AA + )/WT1 was calculated. In the meanwhile, the clinical data from each patient were well documented. Results: The expression levels of WT1 gene (WT1/ABL) in K562, SHI, NB4, BN4/WT1A, and NB4/WT1D cell lines were 2 to 3 magnitudes higher than that in Jurkat and U937 cell lines. The ratios of WT1(17AA+)/WT1 in K562, SHI, Jurkat, U937, NB4, NB4/WT1A and NB4/WT1D were 0. 51, 0. 41, 0. 25, 0. 12, 0. 53, 0. 19 and 0. 62, respectively, suggesting a dominant expression of WT1 (17AA+) isoform in K562 and NB4 cells. The median expression level of WTl gene (WT1/ABL) in 79 newly diagnosed acute leukemia patients was 1. 02, ranging from 0. 03 to 34. 17, which was statistically higher than that in 23 non leukemia controls with median WT1N expression level of 0. 0017(P<0. 001). Furthermore the median ratio of WT1(17AA+ )/WT1 in 79 newly diagnosed acute leukemia patients was 0. 64, ranging from 0. 30 to 0. 93, and there was no statistical difference in the ratios of WT1(17AA+)/WT1 among different subtypes of acute leukemia (F = 0. 152, P = 0. 979). Conclusion: The WT1 gene is expressed in leukemia cell lines and primary leukemia cells with a significantly higher level of the WT1(17AA + ) isoform, and there were no statistical difference in the ratios of WT1(17AA+)/WT1 among different subtypes of acute leukemia.更多还原