FISH法动态观察慢性粒细胞白血病移植治疗后早期Ph阳性细胞  

Dynamic monitering of Ph positive cells during early post-transplant period in chronic granulocytic leukemia by interphase FISH

作　　者：刘世和[1] 李强[1] 韩明哲[1] 庞文新[1] 薄丽津[1] 秦爽[1] 刘旭平[1] 李筱梅[1] 钱林生[1] 

机构地区：[1]中国医学科学院,中国协和医科大学血液学研究所,血液病医院,天津300020

出　　处：《中华血液学杂志》2001年第2期68-71,共4页Chinese Journal of Hematology

基　　金：天津市青年基金!资助项目 (99370 2 5 11)

摘　　要：目的　将间期荧光原位杂交 (FISH)技术应用于慢性粒细胞白血病 (CGL)移植后疗效判断及体内残留Ph阳性细胞的检测。方法　应用FISH、细胞遗传学技术、RT PCR技术对 10名正常人、7例未治疗CGL患者和 11例移植后CGL患者体内Ph阳性细胞进行检测。结果　对 11例CGL患者移植后 1周～ 6年不同时间的 40个样本的实验研究发现 :①移植后 1个月内FISH及RT PCR技术较之常规染色体检查更易检测出Ph阳性的样本 ,但 6个月后RT PCR更敏感 ;②残留Ph阳性细胞随移植后时间延长呈线性递减 ,转阴的中位时间为移植后第 5 7天 (2 2～ 170天 ) ;③ 1例复发患者移植后 1个月Ph阳性残留细胞为 33.6 5 % ,半年后复发 ,而染色体检查于移植后 6个月方发现异常 ,RT PCR检测结果持续阳性 ,无法观测其变化。结论　间期FISH检测比染色体核型、RT PCR更能准确地评价移植后体内残留的白血病细胞的负荷量 ;采用FISH法动态观察残留白血病细胞的变化 ,可以判断疗效 ,发现早期复发 ,及时进行干预性治疗。Objective To assess the effectiveness of transplantation and observe the kinetics of residual leukemic cells during early post transplantation period in chronic granulocytic leukemia(CGL). Methods Ten normal controls, seven untreated CGL patients and eleven CGL patients after allogeneic bone marrow transplantation (allo BMT) or peripheral blood stem cell transplantation (PBSCT) were studied by interphase fluorescence in situ hybridization (I FISH) with the FITC fluorescein labeled BCR probe. Conventional cytogenetic analysis (CCA) and reverse transcriptase polymerase chain reaction (RT PCR) were also employed. Results The normal cutoff was defined as less than 6.68% (±3s) from the analysis of normal controls. The reproduciblity of FISH method was confirmed by serial dilutions by mixing Ph positive cells and normal cells. Forty specimens from 11 CGL patients at 1 week to 6 years post BMT were investigated after PBSCT and the results showed that ①FISH and RT PCR could accurately evaluate Ph(+) cells during early transplantation period (within one month),but RT PCR was more sensitive in six months after BMT. ② residual Ph(+) cells tended to be linear decreasing with the time after transplantation and fell in normal range in most samples three months later. The mean time of normaligation was 57 days (20 to 170 days).③one case relapsed and received a second transplantation. FISH presented more precise data for predicting relapse as compared to CCA and RT PCR. Conclusions I FISH as a useful tool can not only offer more precise assessment of residual leukemic cells compared with CCA and RT PCR, but also provide dynamic monitering during early post BMT period to evaluate the transplantation outcome and predict relapse.

关 键 词：慢性粒细胞白血病 荧光原位杂交 细胞遗传学技术 治疗 

分 类 号：R733.72[医药卫生—肿瘤] R730.5[医药卫生—临床医学]