Pachmann Katharina, Clement Joachim H, Schneider Claus-Peter, Willen Babette, Camara Oumar, Pachmann Ulrich, Höffken Klaus
Abteilung für Experimentalle Hämatologie und Onkologie der Klinik für Innere Medizin II, Friedrich Schiller Universität Jena, Jena, Germany.
Clin Chem Lab Med. 2005;43(6):617-27. doi: 10.1515/CCLM.2005.107.
Detection and quantitation of circulating tumor cells from solid epithelial tumors could become a valuable tool for therapy monitoring if the procedure can be standardized. In the present work we assessed the influence of pre-analytical handling, storage and white blood cell isolation on analysis of a population of spiked tumor cell-line cells and intrinsically present epithelial cells in the peripheral blood of breast and lung cancer patients and the sensitivity of their detection. Sucrose density separation did not enrich epithelial cells, and even depleted them, leading to a gross underestimation of their numbers (3/13 positive, between 2.9 and 50 cells/mL) in comparison to red blood cell lysis (13/13 positive, between 77,200 and 800 cells/mL). Short-term storage of whole blood samples for up to 7 days had little influence on the number of epithelial cells recovered. The effectiveness of magnetic bead enrichment was dependent on the number of relevant cells and the volume used for enrichment. Red blood cell lysis and fluorochrome-labeled antibody staining in a no-wash procedure with subsequent laser scanning cytometry allowed the detection of circulating epithelial cells in 92% of breast and lung cancer patients. Two examples of how this method can be applied for the longitudinal analysis in individual patients are shown, with an increase in numbers preceding relapse and a decrease paralleling tumor reduction. The proposed simple and easy method allows close monitoring, which may help in real-time analysis of the response of solid tumors, especially their systemic component, to therapy and hopefully will contribute to more individually tailored therapy.
如果能实现标准化，那么从实体上皮肿瘤中检测和定量循环肿瘤细胞可能会成为一种有价值的治疗监测工具。在本研究中，我们评估了分析前处理、储存以及白细胞分离对乳腺癌和肺癌患者外周血中添加的肿瘤细胞系细胞群体以及固有存在的上皮细胞分析的影响，以及检测的敏感性。蔗糖密度分离并未富集上皮细胞，甚至使其减少，导致与红细胞裂解相比，上皮细胞数量被严重低估（3/13呈阳性，每毫升2.9至50个细胞）（13/13呈阳性，每毫升77200至800个细胞）。全血样本短期储存长达7天对回收的上皮细胞数量影响不大。磁珠富集的有效性取决于相关细胞的数量和用于富集的体积。在不洗涤程序中进行红细胞裂解和荧光染料标记抗体染色，随后进行激光扫描细胞术，可在92%的乳腺癌和肺癌患者中检测到循环上皮细胞。展示了该方法应用于个体患者纵向分析的两个实例，复发前细胞数量增加，肿瘤缩小同时细胞数量减少。所提出的简单易行的方法能够进行密切监测，这可能有助于实时分析实体肿瘤，尤其是其全身成分对治疗的反应，并有望有助于制定更具个性化的治疗方案。
