双色荧光原位杂交用于慢性粒细胞白血病干扰素治疗效果的监测

Monitoring of the Therapeutic Response of Interferon to Chronic Myeloid Leukemia by Duel-Color Fluorescence in Situ Hybridization

为了探讨双色荧光原位杂交 (D FISH)方法对CML病人干扰素治疗后疗效监测的意义 ,本研究采用D FISH方法对慢性粒细胞白血病 (CML)病人应用干扰素治疗前后骨髓间期核细胞进行bcr abl融合基因的检测 ,并与RT PCR方法检测bcr ablmRNA及传统细胞遗传学方法检测Ph染色体的结果进行比较。结果显示 ,2 2例CML病人D FISHbcr abl融合基因在干扰素治疗前均为阳性 ,其平均检出率在干扰素治疗前后分别为 96 4 %和 5 8.6 % ,其中 2例治疗前Ph染色体阴性病人 ,D FISHbcr abl融合基因在干扰素治疗前后平均检出率分别为 94 .0 %和 70 .1%。D FISH方法检测的 2 2例病人中 ,4例为部分反应 ,4例为微小反应 ,其余 14例为无反应。与传统的细胞遗传学方法相比较具有良好的相关性。结论 :D FISH方法直接检测CML病人间期核细胞的bcr abl融合基因 ,克服了传统的细胞遗传学只能进行分裂中期核细胞分析及RT PCR方法不能定量检测的不足 ,为CML干扰素治疗后克隆缓解程度的评定提供了更方便。

To investigate the significance of duel color fluorescence in situ hybridization (D FISH) in monitoring the response to interferon alpha (IFN α) therapy in patients with chronic myeloid leukemia (CML), the D FISH method was employed to detect the proportion of the interphase nuclei cells with bcr/abl fusion gene in the bone marrow of patients with CML before and after IFN α therapy, and the results were compared with those of bcr/abl fusion mRNA by RT PCR and Philadephia chromsome (Ph) by conventional cytogenetic analysis. The results showed that the mean detectable rate of bcr/abl fusion gene before and after IFN α therapy was 96.4% and 58.6% respectively, in 22 patients who were bcr/abl positive before IFN α therapy by D FISH method, was 94.0% and 70.1% respectively, in 2 patients of Ph negative before treatment. Major, minor and no responses were seen respectively in 4, 4 and 14 cases from 22 patients by D FISH method. The results also showed a good correlation with the analysis of RT PCR and conventional cytogenetics. In conclusion, D FISH method could directly detect the bcr/abl fusion gene of the interphase cells in bone marrow of patients with CML. It can overcome the defect of conventional cytogenetic methods which analyze only the cells in metaphase and the drawback of RT PCR unable to quantify the bcr/abl fusion gene. D FISH provides a more convenient and reliable method for evaluating the degree of clone remission to patients with CML after IFN α therapy.