Flow-FISH同时检测白血病患者细胞端粒长度及表面分化抗原的研究(英文)

Simultaneous Analysis of Telomere Length and Cell Surface Antigen in Leukemia by Multicolor Flow-FISH

本研究应用流式荧光原位杂交(Flow-FISH)方法检测白血病细胞端粒长度,并同时检测表面分化抗原,寻找一个可广泛应用的监测白血病微小残留病的新方法。提取白血病患者骨髓及健康对照者外周血单个核细胞进行Flow-FISH;比较白血病患者与对照者端粒长度的差异;初步分析端粒长度与白血病临床特征、预后的关系;追踪部分患者,观察不同病程中端粒长度与表面分化抗原的变化。结果表明:在34例初发白血病患者中,除CML-CP患者之外,其余端粒长度均较对照组显著缩短。在20例对照者中,端粒长度随年龄增长而缩短,女性较男性端粒长,而初发白血病患者的端粒长度与年龄、性别不相关。在18例AML患者中,可观察到M1患者端粒较长,M5患者端粒较短以及染色体核型高危组患者端粒较低危组患者端粒短的趋势。在可随访的23例急性白血病患者中,端粒长度大于中位数者初治缓解率较高。在5例CML-CP患者中,端粒长度较短者易早期发生急变。14例同时检测表面分化抗原的初发急性白血病患者,缓解后相关抗原表达率均下降,同时端粒延长,其中5例动态追踪的患者中,2例长期缓解者端粒长度在缓解期始终处于正常水平,且无特异抗原表达增高。3例复发者复发后端粒再次缩短,同时特异抗原表达增高,其中2例患者复发时相关抗原阳性细胞群端粒的缩短先于整个细胞群端粒长度的变化,先于骨髓细胞形态学改变。结论:Flow-FISH检测的细胞端粒长度是监测各类白血病疾病状态、评估预后的具有较普遍意义的指标。Flow-FISH同时检测细胞端粒长度和表面分化抗原有利于更早发现异常克隆预测复发,可能可作为监测白血病微小残留病的可广泛应用的新方法。

This study was purposed to explore the feasibility of simultaneous analysis of telomere length and cell surface antigen by multicolor Flow-FISH to assess minimal residual disease （MRD） in leukemia. The telomere length in 34 leukemia patients versus 20 normal controls was compared by using Flow-FISH, and the relationship between telomere length and therapeutic effect and prognosis was analyzed preliminarily. As for those patients with follow-up samples, the changes of telomere length combined with surface antigen in different courses of disease were observed by multicolor Flow-FISH. The results indicated that the telomere length of de novo patients was significantly shorter than that of controls except the patients in chronic myeloid leukemia-chronic phase （CML-CP）. The shorter telomere, the lower complete remission （CR） rates were observed in acute leukemia cases and the shorter duration of CP before onset of blast phase （BP） occured in CML cases. The acute leukemia patients showed longer telomere and fewer cells expressed the related antigen after CR. The telomere length of cases with continued CR remained at normal level during remission, and there was no increased expression of the specific antigen. However, the telomere of relapsed cases shortened again after relapse with elevated specific antigen expression. In the relapsed cases, the telomere of related antigen positive cells shortened ahead of telomere length change of the whole cells and morphologic change of bone morrow cells. It is concluded that analysis of telomere length by flow-FISH manifests the significance for monitoring disease conditions, estimating prognosis and guiding therapy in all kinds of leukemia. The simultaneous analysis of telomere length and cell surface antigen by multicolor flow-FISH may monitor abnormal clone or clonal evolution to predict recurrence more sensitively and specifically, and may provide a promissing and widely applicable method for monitoring MRD in leukemia.