Detection of circulating tumor cells in breast cancer may improve through enrichment with anti-CD146
Detection of circulating tumor cells in breast cancer may improve through enrichment with anti-CD146
复制标题
DOI：
10.1007/s10549-010-0879-y
复制
发表时间：
2011-05-01
影响因子：
3.8
通讯作者：
Sleijfer, Stefan
中科院分区：
文献类型：
--
作者：
Mostert, Bianca;Kraan, Jaco;Sleijfer, Stefan
大多数检测循环肿瘤细胞 (CTC) 的检测依赖于肿瘤细胞上的 EpCAM 表达。最近，我们小组报道，与其他分子乳腺癌亚型相比，“正常样”细胞系缺乏 EpCAM 表达，因此当使用基于 EpCAM 的技术捕获 CTC 时会被遗漏 [J Natl Cancer Inst 101(1):61-66, 2009]。这里引入使用CD146来检测EpCAM阴性CTC，从而改进CTC检测。在我们的 41 个乳腺癌细胞系组中评估了 CD146 和 EpCAM 表达。来自 14 个细胞系的细胞（其中 9 个与正常细胞系相似）被掺入健康供体血液中。使用 CellSearch (TM) 技术，通过添加负载 EpCAM 和/或 CD146 抗体的铁磁流体，然后对细胞角蛋白和 DAPI 进行染色，对 7.5 ml 全血进行 CTC 富集。造血细胞和循环内皮细胞 (CEC) 分别用 CD45 和 CD34 复染。类似的方法也适用于 20 名晚期乳腺癌患者的血液样本。 9 个正常乳腺癌细胞系中有 8 个缺乏 EpCAM 表达，但表达 CD146。这 8 个中的 5 个可以通过抗 CD146 铁磁流体充分回收。在 20 名晚期乳腺癌患者中，用抗 EpCAM 和抗 CD146 铁磁流体对 CTC 进行计数，其中 9 名患者具有 CD146+ CTC。来自缺乏 EpCAM 表达的乳腺癌细胞系的细胞经常表达 CD146，并且可以通过抗 CD146 铁磁流体来恢复。 CD146+ CTC 存在于晚期乳腺癌患者的外周血中。联合使用抗 CD146 和抗 EpCAM 可能会改善乳腺癌患者的 CTC 检测。
Most assays to detect circulating tumor cells (CTCs) rely on EpCAM expression on tumor cells. Recently, our group reported that in contrast to other molecular breast cancer subtypes, "normal-like" cell lines lack EpCAM expression and are thus missed when CTCs are captured with EpCAM-based technology [J Natl Cancer Inst 101(1):61-66, 2009]. Here, the use of CD146 is introduced to detect EpCAM-negative CTCs, thereby improving CTC detection. CD146 and EpCAM expression were assessed in our panel of 41 breast cancer cell lines. Cells from 14 cell lines, 9 of which normal-like, were spiked into healthy donor blood. Using CellSearch (TM) technology, 7.5 ml whole blood was enriched for CTCs by adding ferrofluids loaded with antibodies against EpCAM and/or CD146 followed by staining for Cytokeratin and DAPI. Hematopoietic cells and circulating endothelial cells (CECs) were counterstained with CD45 and CD34, respectively. A similar approach was applied for blood samples of 20 advanced breast cancer patients. Eight of 9 normal-like breast cancer cell lines lacked EpCAM expression but did express CD146. Five of these 8 could be adequately recovered by anti-CD146 ferrofluids. Of 20 advanced breast cancer patients whose CTCs were enumerated with anti-EpCAM and anti-CD146 ferrofluids, 9 had CD146+ CTCs. Cells from breast cancer cell lines that lack EpCAM expression frequently express CD146 and can be recovered by anti-CD146 ferrofluids. CD146+ CTCs are present in the peripheral blood of breast cancer patients with advanced disease. Combined use of anti-CD146 and anti-EpCAM is likely to improve CTC detection in breast cancer patients.