Heterogeneous detection of circulating tumor cells in patients with colorectal cancer by immunomagnetic enrichment using different EpCAM-specific antibodies.
Heterogeneous detection of circulating tumor cells in patients with colorectal cancer by immunomagnetic enrichment using different EpCAM-specific antibodies.
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DOI：
10.1186/1472-6750-10-35
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发表时间：
2010-04-28
影响因子：
3.5
通讯作者：
Koch M
中科院分区：
文献类型：
--
作者：
Antolovic D;Galindo L;Carstens A;Rahbari N;Büchler MW;Weitz J;Koch M
循环肿瘤细胞（CTC）和播散性肿瘤细胞（DTC）被认为是结直肠癌转移的主要原因，因此CTC的检测可以作为结直肠癌患者个体化预后的指标。因此，已经开发了一系列的免疫磁性富集CTC的方法，使用各种针对上皮细胞粘附分子（EpCAM）的单克隆抗体。然而，尚不清楚是否所有市售的EpCAM抗体都显示出相同的灵敏度和特异性。此外，目前还不清楚哪种样品制备和细胞提取方法最适合于免疫磁性富集和检测CTC。在这项研究中，我们的目的是调查是否检测CTC的细胞角蛋白20逆转录酶-聚合酶链反应（CK 20 RT-PCR）可能会受到影响，使用各种上皮细胞粘附分子（EpCAM）抗体免疫磁性分离CTC。使用两种EpCAM抗体（mAb BerEP 4和mAb KS 1/4）在39名结肠直肠癌患者的血液样品中进行免疫磁性富集，我们在每个患者中发现关于肿瘤细胞检测的异质性结果。在全血样本的肿瘤细胞加标实验中，与预包被mAb BerEP 4 Dynabeads相比，使用mAb KS 1/4包被的免疫磁珠进行CK 20 RT-PCR测定的灵敏度更高。在免疫磁性富集之前用Ficoll梯度离心提取MNC组分导致CK 20 RT-PCR测定的更高灵敏度。我们的结论是，分离和检测CTC的免疫磁性富集方法是非常依赖于所使用的EpCAM克隆。更多患者的进一步研究应阐明富集方案是否影响肿瘤细胞检测方案的预后价值。
Circulating tumor cells (CTC) and disseminated tumor cells (DTC) are thought to be responsible for metastasis, so the detection of CTC may serve as individual prognostic factor in patients suffering from colorectal cancer. Therefore, a series of immunomagnetic enrichment methods for CTC have been developed using a variety of monoclonal antibodies against the Epithelial Cell Adhesion Molecule (EpCAM). However, it remains unclear whether all commercially available EpCAM antibodies show the same sensitivity and specificity. Furthermore, it remains unclear which method of sample preparation and cell extraction is most suitable for immunomagnetic enrichment and detection of CTC. In this study, we aimed to investigate whether the detection of CTC by a cytokeratin 20 reverse transcriptase-polymerase chain reaction (CK20 RT-PCR) may be influenced by the use of various Epithelial Cell Adhesion Molecule (EpCAM) antibodies for immunomagnetic isolation of CTC. Using both EpCAM antibodies (mAb BerEP4 and mAb KS1/4) for immunomagnetic enrichment in blood samples of 39 patients with colorectal cancer we found heterogenous results in each patient with regard to tumor cell detection. In the tumor cell spiking experiments with whole blood samples the sensitivity of the CK 20 RT-PCR assay was higher using immunomagnetic beads coated with mAb KS1/4 compared to precoated mAb BerEP4 Dynabeads. Extraction of MNC fraction with Ficoll gradient centrifugation prior to immunomagnetic enrichment resulted in a higher sensitivity of the CK 20 RT-PCR assay. We concluded that isolation and detection of CTC with immunomagnetic enrichment methods is critically dependent on the used EpCAM clone. Further studies with a larger number of patients should clarify if the enrichment protocol influences the prognostic value of the tumor cell detection protocol.
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影响因子：
11.5
作者：
Willipinski-Stapelfeldt, B;Riethdorf, S;Pantel, K
通讯作者：
Pantel, K
影响因子：
6.4
作者：
Koch, Moritz;Kienle, Peter;Weitz, Jurgen
通讯作者：
Weitz, Jurgen
影响因子：
--
作者：
Vlems, F;Soong, R;van Muijen, G
通讯作者：
van Muijen, G
影响因子：
45.3
作者：
Cohen, Steven J.;Punt, Cornelis J. A.;Meropol, Neal J.
通讯作者：
Meropol, Neal J.
影响因子：
4.7
作者：
Guo, JM;Xiao, BX;Liu, Z
通讯作者：
Liu, Z