DSP30和IL-2在慢性淋巴细胞白血病常规染色体检测中的应用

Chromosomal aberrations detection in chronic lymphocytic leukemia by conventional cytogenetics using DSP30 and IL-2

目的研究未甲基化胞嘧啶鸟嘌呤二核苷酸寡脱氧核苷酸(DSP30)和IL-2在慢性淋巴细胞白血病(CLL)常规染色体检测中的价值。方法DSP30联合IL-2刺激CLL细胞增殖,CLL细胞培养72 h后进行染色体制备,采用R显带法进行核型分析。对85例患者进行FISH检测,与同期核型结果进行比较。结果89例CLL患者中,无分裂象者5例,84例(94.38%)患者染色体分析成功,51例检测出染色体异常,异常检出率为60.71%(51/84),复杂核型者占52.94%(27/51)。对85例CLL患者进行了FISH检测,FISH探针为D13S25、RB1、P53、ATM、cen12。FISH检测结果显示74例异常,异常检出率为87.06%(74/85),其中复杂核型2例,占2.70%(2/74)。85例进行FISH检测的CLL患者中,50例常规染色体核型分析异常,30例核型正常,5例无分裂象,异常检出率为62.50%(50/80);其中FISH检测异常而核型正常者25例,核型异常而FISH检测正常者4例,两者结合检测出异常者78例,异常检出率91.76%。FISH检测到的13q-异常率明显高于常规核型分析(69.41%对16.67%,P<0.001),11q-、+12、17p-异常检出率两种方法的差异无统计学意义(P>0.05)。常规核型分析对复杂异常的检出率(50.98%)高于FISH(2.70%)。另外,根据FISH结果进行预后分层的11例低危和9例中危患者均为复杂核型,根据核型结果被归为高危组。结论DSP30联合IL-2可以提高CLL患者常规染色体异常的检出率,对复杂异常核型的检测更为有效。FISH异常检出率高于常规核型分析方法,对缺失片段较小的13q-异常更为敏感,两者结合不仅将染色体异常检出率提高至91.76%,还可对CLL患者进行更为准确的预后分层,为临床提供更多信息。

Objective To study the value of unmethylated cytosine guanine dinucleotide oligodeoxynucleotide(DSP30)and IL-2 in the conventional cytogenetic(CA)detection of the chromosomal aberrations in chronic lymphocytic leukemia(CLL).Methods Bone marrow or peripheral blood cells of CLL patients were cultured with DSP30 plus IL-2 for 72 h,following which R-banding analysis was conducted.Fluorescence in situ hybridization(FISH)was performed in 85 patients.CA results were compared with data obtained by FISH.Results Among 89 CLL patients,the success rate of chromosome analysis was 94.38%(84/89).Clonal aberrations were detected in 51 patients(51/84,60.71%).Of them,27(27/51,52.94%)were complex karyotype.Among 85 CLL patients tested by FISH,chromosomal abnormalities were detected in 74(74/85,87.06%)patients,of which 2(2/74)patients were complex karyotypes,accounting for 2.70%.Of the 85 CLL patients examined by FISH,50 had abnormal karyotype analysis,30 had normal karyotype,5 failed to have chromosome analysis.Among them,25 cases showed clonal aberrations by FISH assay but normal by CA,and 4 cases were normal by FISH but displayed aberrations in chromosome analysis,and totally 78(91.76%)cases with abnormality detected by the combination of the two methods.The frequency of 13q-abnormality detected by FISH was significantly higher than that by CA analysis(69.41%vs 16.67%,P<0.001),while the frequency of 11q-,+12 and 17p-detected by two methods showed no significant difference(P>0.05).The detection rate of complex abnormalities in conventional karyotype analysis was higher than that in FISH(50.98%vs 2.70%).In addition,11 low-risk and 9 intermediate-risk patients according to FISH results showed complex karyotype by cytogenetics,and were classified into high-risk cytogenetic subgroup.Conclusion DSP30 and IL-2 are effective in improving the detection rate of CA in CLL patients(60.71%)and CA is more effective to detect complex karyotype.However,FISH had a higher overall abnormality detection rate(87.06%)than CA,especially for 13q-.The combination of CA and FISH not only enhanced the detection rate of clonal aberrations to 91.76%,but also provided more precise prognosis stratification for CLL patients,thus to provide more information for clinical implication.