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目的探讨慢性粒细胞白血病(CML)患者骨髓细胞中 Wilms 瘤抑癌基因(WT1)的表达水平及其临床意义。方法建立实时定量 RT-PCR 方法,采用 Light Cycler PCR 仪检测了46例(109份骨髓细胞 cDNA 标本)CML 患者和23例非白血病患者骨髓细胞中 WT1及内参β胆色原脱氢酶(GAPDH)的表达水平,以 WT1_N=(WT1拷贝数/GAPDH 拷贝数)×10~4计算 WT1表达水平。结果23份 CML 加速期与22份 CML 急变期患者骨髓细胞中 WT1_N 的中位表达水平分别为103.71和129.44,明显高于64份 CML 慢性期和对照组(分别为3.44和1.47,P<0.01),对照组与 CML 慢性期之间 WT1基因表达差异无统计学意义;CML 加速期与急变期之间 WT1基因表达差异也无统计学意义(P<0.05)。WT1基因表达水平与 BCR/ABL 融合基因表达水平具有一定的相关性。对其中7例 CML 患者行异基因骨髓移植前后动态检测 WT1上升可提示白血病复发。结论 CML 患者加速急变期骨髓细胞中 WT1基因表达升高,具有参考意义。
Objective To investigate the expression of Wilms tumor suppressor gene (WT1) in bone marrow cells of patients with chronic myelogenous leukemia (CML) and its clinical significance. Methods Real-time quantitative RT-PCR was used to detect the expression of WT1 and GβDH in bone marrow cells of 46 patients (109 bone marrow cell cDNA samples) and 23 non-leukemia patients by Light Cycler PCR. WT1_N = (WT1 copy number / GAPDH copy number) × 10-4 was used to calculate the expression level of WT1. Results The median expression levels of WT1_N in 23 CML accelerated phase and 22 CML patients were 103.71 and 129.44, respectively, which were significantly higher than those in 64 CML chronic phase and control group (3.44 and 1.47 respectively, P <0.01) There was no significant difference in the expression of WT1 between the control group and the chronic phase of CML. There was also no significant difference in WT1 gene expression between the accelerated and abrupt CML phases (P <0.05). WT1 gene expression level and BCR / ABL fusion gene expression level has a certain correlation. The dynamic detection of WT1 in 7 CML patients before and after allogeneic bone marrow transplantation may indicate the relapse of leukemia. Conclusions The expression of WT1 gene in bone marrow cells of patients with CML during accelerated blastic phase is elevated, which is of reference value.