多发性骨髓瘤患者81例细胞及分子遗传学研究

目的 探讨多发性骨髓瘤(MM)患者遗传学特征,评价添加细胞因子及延长培养时间对MM染色体核型分析的影响和间期荧光原位杂交(FISH)检测RB1与P53基因缺失的意义.方法 采用骨髓24h短期培养和G显带技术对81例MM患者进行染色体核型分析,其中28例患者同时采用添加IL-6(终浓度10 μg/L)和重组粒-巨噬细胞集落刺激因子(GM-CSF,终浓度40 μg/L)的6d培养法后的G显带核型分析,31例患者采用间期FISH方法检测RB1与P53基因缺失.结果 81例患者中有75例患者有足够可供分析的中期分裂象,其中31例(41.3％)检出异常克隆.在31例检出染色体异常核型的患者中,单纯染色体数目异常4例(12.9％),单纯染色体结构异常11例(35.5％),染色体数目和结构异常同时存在16例(51.6％).28例患者同时采用两种培养方法对照研究结果显示,骨髓24h短期培养法异常克隆检出率为25.0％,添加细胞因子的6d延期培养法异常克隆检出率为51.9％ (P =0.026).31例患者间期FISH结果显示RB1基因缺失10例(32.3％),P53基因缺失11例(35.5％),5例患者RBI和P53基因均缺失.结论 MM染色体核型异常半数以上为染色体数目和结构异常同时存在,显带技术是遗传学检测的基础,添加细胞因子的长期培养法可提高显带技术的异常克隆检出率,间期FISH是一种检测MM患者骨髓瘤细胞基因缺失的敏感方法,具有临床应用价值.

The cytogenetic and molecular genetic study of 81 multiple myeloma patients

Objective To explore the cytogenetic characteristics of multiple myeloma (MM) patients,to evaluate the effect of a long-term culture stimulated by cytokines on cytogenetic study of MM,and to investigate the clinical detection value of RB1 and P53 deletion in interphase plasma cells by using fluorescence in situ hybridization (FISH).Methods Karyotype analysis was performed in 81 MM patients by using the short-term culture of bone marrow cell and G-banding technique.Among the 81 MM patients,28 patients used two culture methods:one was the short-term culture and the other was to culture cells for 6 days with recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) (40 μg/L) and IL-6 (10 μg/L).RB1 and P53 deletion were detected on interphase plasma cells by using FISH in 31 patients.Results Among the 81 patients,75 had enough metaphases for analysis.Among the 75 patients,31 (41.3％) had clonal karyotypic abnormalities including 4 numeric abnormalities,11 structural abnormalities and 16 both abnormalities.Among the 28 patients using two culture methods,the clonal karyotypic abnormalities were detected in 6 patients(25.0％ ) in the group of cultured for 24 hours,and 14 patients (51.9％) in 6-day culture group with a significant difference (P =0.026).RB1 deletion and P53 deletion were detected in 10 patients (32.3％ ) and 11 patients(35.5％ ),respectively,with both RB1 and P53 deletions be detected in 5 patients ( 16.1％ ).Conclusions More than half of the tested MM patients have both numeric and structural chromosome abnormalities.The karyotype analysis using banding technique is basic cytogenetic study.Extended culture in the presence of IL-6 and GM-CSF could improve the efficiency of cytogenetic analysis to MM.Interphase FISH is a sensitive method of clinical application significance to detect the gene deletion of MM.