Novak Urban, Oppliger Leibundgut Elisabeth, Hager Jörg, Mühlematter Dominique, Jotterand Martine, Besse Celine, Leupin Nicolas, Ratschiller Daniel, Papp Jeanette, Kearsey Gina, Aebi Stefan, Graber Hans, Jaggi Rolf, Lüthi Jean-Marc, Meyer-Monard Sandrine, Lathrop Mark, Tobler Andreas, Fey Martin F
Department of Clinical Research and Medical Oncology/Hematology, Inselspital (University Hospital), Bern, Switzerland.
Blood. 2002 Sep 1;100(5):1787-94.
The most frequent chromosomal aberrations in B-cell chronic lymphocytic leukemia (B-CLL) are deletions on 13q, 11q, and 17p, and trisomy 12, all of which are of prognostic significance. Conventional cytogenetic analysis and fluorescence in situ hybridization (FISH) are used for their detection, but cytogenetic analysis is hampered by the low mitotic index of B-CLL cells, and FISH depends on accurate information about candidate regions. We used a set of 400 highly informative microsatellite markers covering all chromosomal arms (allelotyping) and automated polymerase chain reaction (PCR) protocols to screen 46 patients with typical B-CLL for chromosomal aberrations. For validation, we compared data with our conventional karyotype results and fine mapping with conventional single-site PCR. All clonal cytogenetic abnormalities potentially detectable by our microsatellite PCR (eg, del13q14 and trisomy 12) were picked up. Allelotyping revealed additional complex aberrations in patients with both normal and abnormal B-CLL karyotypes. Aberrations detectable in the samples with our microsatellite panel were found on almost all chromosomal arms. We detected new aberrant loci in typical B-CLL, such as allelic losses on 1q, 9q, and 22q in up to 25% of our patients, and allelic imbalances mirroring chromosomal duplications, amplifications, or aneuploidies on 2q, 10p, and 22q in up to 27% of our patients. We conclude that allelotyping with our battery of informative microsatellites is suitable for molecular screening of B-CLL. The technique is well suited for analyses in clinical trials, it provides a comprehensive view of genetic alterations, and it may identify new loci with candidate genes relevant in the molecular biology of B-CLL.
B 细胞慢性淋巴细胞白血病（B-CLL）中最常见的染色体畸变是 13q、11q 和 17p 缺失以及三体 12，所有这些都具有预后意义。传统的细胞遗传学分析和荧光原位杂交（FISH）用于检测这些畸变，但细胞遗传学分析因 B-CLL 细胞的低有丝分裂指数而受到阻碍，FISH 则依赖于有关候选区域的准确信息。我们使用一组覆盖所有染色体臂的 400 个高信息量微卫星标记（等位基因分型）和自动化聚合酶链反应（PCR）方案，对 46 例典型 B-CLL 患者进行染色体畸变筛查。为了进行验证，我们将数据与我们的传统核型分析结果进行比较，并用传统的单位点 PCR 进行精细定位。我们的微卫星 PCR 可能检测到的所有克隆性细胞遗传学异常（例如，del13q14 和三体 12）均被检出。等位基因分型揭示了 B-CLL 核型正常和异常患者中额外的复杂畸变。我们的微卫星检测板在几乎所有染色体臂上都发现了样本中可检测到的畸变。我们在典型 B-CLL 中检测到新的异常位点，例如高达 25%的患者中 1q、9q 和 22q 上的等位基因缺失，以及高达 27%的患者中 2q、10p 和 22q 上反映染色体重复、扩增或非整倍体的等位基因失衡。我们得出结论，使用我们这一系列信息丰富的微卫星进行等位基因分型适用于 B-CLL 的分子筛查。该技术非常适合临床试验分析，它提供了遗传改变的全面视图，并且可能识别出与 B-CLL 分子生物学相关的具有候选基因的新位点。
