Wiedswang G, Borgen E, Karesen R, Naume B
Department of Surgery Ullevål University Hospital Oslo, Oslo, Norway.
Cytotherapy. 2003;5(1):40-5. doi: 10.1080/14653240310000065.
The aim of this study was to determine the influence and significance of different aspiration sites and the number of mononuclear cells (MNC) analyzed on the frequency of isolated tumor cell (ITC) detection by immunocytochemistry (ICC) in BM aspirates from breast-cancer patients.
BM aspirates were collected from the two anterior and two posterior crests just prior to primary surgery. The BM was processed separately from the anterior and the posterior crests, and cytospins (2 x 10(6) MNC) were prepared for ICC examination. The remaining cells were pooled, followed by cytospin preparation and ICC analysis (2 x 10(6) MNC/test). In addition, a fraction of the pooled cells were further processed by negative immunomagnetic selection, for enrichment of ITC. Out of 100 patients selected, 97 were further analyzed.
The ICC examination from the separate crests revealed 37 positive BMs from the anterior iliac crest and 30 positive from the posterior crest. Twenty-one of the samples were positive at both sides. Five patients had 10 or more ITCs detected. In these, an unequal distribution of ITCs between the sides was observed, but in favor of neither. ICC analysis of 2, 4 and 6 x 10(6) MNC revealed respectively 22, 46 and 52 positive BMs out of the 97 analyzed. These results were correlated to the clinical outcome after a median 43 months follow-up. Thirteen of the patients underwent systemic relapse. Analyzing 2 x 10(6) MNC by ICC, 27.3% of the BM+ patients developed systemic disease, compared with 9.3% of the BM+ patients (P = 0.0056, log rank test). Analyzing 6 x 10(6) MNC reduced the correlation between ITC in BM and clinical outcome.
No significant difference in the detection rate of ITCs from the anterior and the posterior iliac crests was found, although the distribution of ITCs did show a great variability. Analyzing a higher number of BM cells increased the number of positive BM specimens detected. However, this increased detection rate reduces the prediction by ICC of early systemic relapse.
本研究的目的是确定不同取材部位以及分析的单个核细胞（MNC）数量对乳腺癌患者骨髓穿刺液中通过免疫细胞化学（ICC）检测孤立肿瘤细胞（ITC）频率的影响及意义。
在初次手术前从双侧前、后髂嵴采集骨髓穿刺液。分别对前、后髂嵴的骨髓进行处理，并制备细胞涂片（2×10⁶个MNC）用于ICC检查。将剩余细胞合并，随后制备细胞涂片并进行ICC分析（2×10⁶个MNC/检测）。此外，对合并细胞的一部分进行阴性免疫磁选进一步处理，以富集ITC。在入选的100例患者中，97例进行了进一步分析。
对分开的髂嵴进行ICC检查显示，髂前嵴有37例骨髓阳性，髂后嵴有30例阳性。21例样本双侧均为阳性。5例患者检测到10个或更多ITC。在这些患者中，观察到双侧ITC分布不均，但无明显偏向。对97例患者分析2×10⁶、4×10⁶和6×10⁶个MNC的ICC结果分别显示22例、46例和52例骨髓阳性。这些结果与中位随访43个月后的临床结局相关。13例患者发生全身复发。通过ICC分析2×10⁶个MNC，骨髓阳性患者中有27.3%发生全身疾病，而骨髓阳性患者中这一比例为9.3%（P = 0.0056，对数秩检验）。分析6×10⁶个MNC降低了骨髓中ITC与临床结局之间的相关性。
虽然ITC的分布确实存在很大变异性，但在髂前嵴和髂后嵴检测ITC的检出率无显著差异。分析更多数量的骨髓细胞增加了检测到的阳性骨髓标本数量。然而，这种增加的检出率降低了ICC对早期全身复发的预测能力。
