Dong Hiep P, Holth Arild, Berner Aasmund, Davidson Ben, Risberg Björn
Pathology Clinic, Radiumhospitalet-Rikshospitalet Medical Center, University of Oslo, Montebello, N-0310 Oslo, Norway.
Cytometry B Clin Cytom. 2007 Sep;72(5):332-43. doi: 10.1002/cyto.b.20172.
Data regarding the role of flow cytometry (FCM) in the characterization of malignant effusions are limited to date. In the present study, we optimized the conditions for FCM immunphenotyping of effusions using a four-color analysis and investigated aspects related to the advantages and limitations of this method in this setting.
FCM analysis optimization for the study of epithelial cells was undertaken using five carcinoma cell lines, and subsequently applied to malignant pleural and peritoneal effusions using antibodies against epithelial and mesothelial markers (Ber-EP4 and EMA), CD138, and integrin subunits. FCM of frozen versus fresh specimens and the performance of FCM compared to immunhistochemistry were evaluated.
FCM optimization was achieved and applied to clinical specimens, with resulting detection of epithelial markers and adhesion molecules on cancer cells. Frozen clinical specimens and cell lines showed reduced CD138 expression compared to fresh specimens, with conservation of the remaining epitopes. FCM generally showed comparable performance to immunhistochemistry.
FCM is an effective method for characterization of cancer cells in clinical effusion specimens in both the diagnostic and research setting, and is comparable to immunhistochemistry in terms of sensitivity and specificity, with the additional advantage of providing quantitative data. The majority of epitopes are conserved in frozen cells, but a minority may be lost, suggesting that the thorough testing of each antibody in both conditions is mandatory.
迄今为止，关于流式细胞术（FCM）在恶性积液特征分析中作用的数据有限。在本研究中，我们使用四色分析法优化了积液FCM免疫表型分析的条件，并研究了该方法在此背景下的优势和局限性相关的方面。
使用五种癌细胞系对上皮细胞研究进行FCM分析优化，随后使用针对上皮和间皮标志物（Ber-EP4和EMA）、CD138和整合素亚基的抗体应用于恶性胸腔和腹腔积液。评估了冷冻标本与新鲜标本的FCM以及FCM与免疫组织化学相比的性能。
实现了FCM优化并应用于临床标本，从而检测到癌细胞上的上皮标志物和黏附分子。与新鲜标本相比，冷冻临床标本和细胞系显示CD138表达降低，其余表位得以保留。FCM总体表现与免疫组织化学相当。
FCM是在诊断和研究环境中对临床积液标本中的癌细胞进行特征分析的有效方法，在敏感性和特异性方面与免疫组织化学相当，还具有提供定量数据的额外优势。大多数表位在冷冻细胞中得以保留，但少数可能会丢失，这表明在两种条件下对每种抗体进行全面测试是必要的。
