Van Der Lely N, Poddighe P, Wessels J, Hopman A, Geurts van Kessel A, De Witte T
Department of Internal Medicine, University Hospital, Nijmegen, The Netherlands.
Leukemia. 1995 Jul;9(7):1167-72.
Interphase cytogenetics was used to investigate the clonal origin of bone marrow (BM) cells, peripheral blood (PB) cells, and in vitro cultured progenitor cells of five patients with acute myeloid leukemia (AML) and myelodysplasia (MDS). A new in situ hybridization (ISH) technique was used to examine the origin of the progenitor cells. Two patients with respectively, trisomy 8 and polyploidy as ISH marker were studied both at presentation and during remission. At presentation, the in vitro cultured clusters of both cases appeared diploid. Therefore, despite the abnormal growth patterns, the cultured progenitors could have been residual normal cells. Alternatively, they could have originated from a preleukemic clone with a normal karyotype. In both cases abnormal BM and/or PB cells (less than 6%) were detected with ISH during remission, indicating partially or completely clonal remissions in these patients. Both patients have relapsed. One patient with trisomy 10 as ISH marker was analyzed during myelodysplastic phase and after progression to AML. On both occasions, abnormally appearing clusters were cultured. However, only part of the clusters carried trisomy 10. The presence of a subclone characterized by trisomy 10 and an abnormally growing (pre)leukemic clone without trisomy 10 may explain this observation. Monosomy 1 and 17 were respectively used as ISH markers in two other AML patients. All in vitro cultured clusters carried the numerical abnormality. Long-term liquid cultures of these leukemias were performed for 10-20 days. In both cases, no residual normal clonogenic cells could be detected. Therefore, the selective growth advantage of normal progenitor cells in long-term marrow cultures could not be demonstrated in these two patients with leukemia. This paper illustrates the usefulness of ISH to study the biology of AML at the clonogenic level during preleukemic phase, active disease, remission, and under in vitro culture conditions. It is a sensitive technique which allows individual analysis of large numbers of small aggregates and single cells in culture.
采用间期细胞遗传学方法研究了5例急性髓系白血病（AML）和骨髓增生异常综合征（MDS）患者的骨髓（BM）细胞、外周血（PB）细胞及体外培养祖细胞的克隆起源。运用一种新的原位杂交（ISH）技术检测祖细胞的起源。分别以8号染色体三体和多倍体作为ISH标记的2例患者在初诊时和缓解期均进行了研究。初诊时，这2例患者的体外培养集落均显示为二倍体。因此，尽管生长模式异常，但培养的祖细胞可能是残留的正常细胞。或者，它们可能起源于核型正常的白血病前期克隆。2例患者在缓解期经ISH检测均发现异常的BM和/或PB细胞（少于6%），表明这些患者部分或完全获得克隆性缓解。2例患者均已复发。以10号染色体三体作为ISH标记的1例患者在骨髓增生异常阶段及进展为AML后进行了分析。两次检测时均培养出外观异常的集落。然而，只有部分集落携带10号染色体三体。存在以10号染色体三体为特征的亚克隆以及无10号染色体三体的异常生长的（前）白血病克隆可能解释了这一现象。另外2例AML患者分别以1号和17号染色体单体作为ISH标记。所有体外培养的集落均存在数目异常。对这些白血病进行了10 - 20天的长期液体培养。2例患者均未检测到残留的正常克隆形成细胞。因此，在这2例白血病患者中未能证实正常祖细胞在长期骨髓培养中有选择性生长优势。本文阐述了ISH在白血病前期、疾病活动期、缓解期及体外培养条件下研究AML克隆形成水平生物学特性的实用性。这是一种敏感的技术，可对培养中的大量小聚集体和单个细胞进行个体分析。
