Hildebrandt M, Mapara M Y, Körner I J, Bargou R C, Moldenhauer G, Dörken B
Department of Medical Oncology and Tumor Immunology, Robert Rössle-Klinik, Max Delbrück Center for Molecular Medicine, Medical Faculty of the Humboldt University, Berlin, Germany.
Exp Hematol. 1997 Jan;25(1):57-65.
A modified reverse transcriptase-polymerase chain reaction (RT-PCR) technique was established with the aim of monitoring the tumor cell contamination in peripheral blood stem cells harvested from breast cancer patients. In an experimental approach, single cell suspensions of different breast cancer cell lines were mixed to normal peripheral blood mononuclear cells in order to 1) determine the sensitivity of tumor cell detection within PBMC and 2) compare polymerase chain reaction in its capacity of monitoring the efficiency of immunomagnetic purging using the magnetic cell separation (MACS) system to immunocytochemical staining. Several target sequences were assessed for their indicative potential and specificity allowing the detection of breast cancer cells by RT-PCR. Among the sequences evaluated, epithelial growth factor receptor (EGF-R) mRNA and Cytokeratin 19 mRNA were shown to be highly specific and sensitive markers for the detection of breast cancer cells within normal peripheral blood mononuclear cells and for the evaluation of the efficiency in immunomagnetic purging. In addition, we were able to show that the MACS is a potent and efficient tool for the selection of tumor cells from peripheral blood mononuclear cells, thus establishing its value for clinical scale immunomagnetic purging.
建立了一种改良的逆转录聚合酶链反应（RT-PCR）技术，旨在监测从乳腺癌患者采集的外周血干细胞中的肿瘤细胞污染情况。在实验方法中，将不同乳腺癌细胞系的单细胞悬液与正常外周血单核细胞混合，以便：1）确定外周血单核细胞内肿瘤细胞检测的灵敏度；2）比较聚合酶链反应监测使用磁性细胞分离（MACS）系统进行免疫磁珠清除效率的能力与免疫细胞化学染色的能力。评估了几个靶序列的指示潜力和特异性，以通过RT-PCR检测乳腺癌细胞。在所评估的序列中，上皮生长因子受体（EGF-R）mRNA和细胞角蛋白19 mRNA被证明是检测正常外周血单核细胞内乳腺癌细胞以及评估免疫磁珠清除效率的高度特异性和敏感的标志物。此外，我们能够证明MACS是从外周血单核细胞中选择肿瘤细胞的有效工具，从而确立了其在临床规模免疫磁珠清除中的价值。
