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.
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。.
Keywords: Cytogenetics; DSP30; FISH; IL-2; Leukemia, lymphocytic, chronic.