组合探针荧光原位杂交检测多发性骨髓瘤常见染色体异常

Fluorescence in situ hybridization based on a panel of probes for detection of common cytogenetic abnormalities in multiple myeloma

目的:研究应用磁珠分选联合组合探针荧光原位杂交(FISH)检测多发性骨髓瘤(MM)患者常见染色体异常情况。方法:收集MM患者新鲜骨髓标本,采用CD138磁珠分选富集浆细胞,应用组合探针FISH检测MM常见的染色体异常,同一骨髓标本同时进行短期培养染色体核型分析,比较两种方法对染色体异常检出率的差异。结果:应用组合探针FISH检测95例初诊MM患者有72例(75.8%)检出染色体异常克隆,其中42例(44.2%)检出RB1(13q14)缺失,40例(42.1%)检出CKS1B(1q21)扩增,8例(8.4%)检出CDKN2C(1p32)缺失,12例(12.6%)检出TP53缺失,IGH/CCND1、IGH/FGFR3检出率均为14.7%(14/95),未检出IGH/MAF融合基因。染色体核型分析有32例(33.7%)检出染色体异常克隆,应用组合探针FISH的染色体异常检出率高于染色体核型分析(75.8%vs.33.7%,P=0.000)。结论:初诊MM患者应采用组合探针FISH检测经分选富集浆细胞的染色体异常,有利于提高染色体异常检出率,对MM患者的危险度分层、预后判断具有重要意义。

Objective To evaluate the advantages of plasma cell enrichment combined with fluorescence in situ hybridization （FISH） based on a panel of probes by the conventional cytogenetic （CC） analysis. Methods Fresh heparinized bone marrow samples were collected by bone marrow biopsy. Plasma cells were enriched in BM sampies using a magnetic cell-sorting procedure to select CD138＋ cells. The common chromosome abnormalities of MM were detected by FISH based on a panel of probes and CC analysis after short-term culture of the BM cells, in order to compare the differences between these two methods for the frequency of common cytogenetic abnormalities. Results 72 of 95 （75.8%） MM patients were found to carry clonal chromosome abnormalities by FISH. And RB1 deletion was the highest at 44.2% （42/95） followed by CKS1B （I@1） amplification （42.1%）. The frequencies of CD- KN2C （lp32） deletion, TP53 deletion, IGH/CCND1 and IGH/FGFR3 were 8.4% （8/95）, 12.6% （12/95）, 14.7% （14/ 95） and 14.7% （14/95）, respectively. IGH/MAF was negative. Thirty-two of 95 （33.7%） patients were found to carry clonal aberrations by CC analysis. The frequency of chromosome abnormalities detected by FISH was significantly higher than CC analysis （75.8% vs 33.7%, P = 0.000）. Conclusion Plasma cell enrichment combined with FISH based on a panel of probes can greatly increase the frequency of chromosome abnormalities, which provides cytogenetic basis for risk stratification and prognosis of MM patients.