Heterogeneous atypical cell populations are present in blood of metastatic breast cancer patients.
Heterogeneous atypical cell populations are present in blood of metastatic breast cancer patients.
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DOI：
10.1186/bcr3622
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发表时间：
2014-03-06
期刊：
影响因子：
--
通讯作者：
Chalmers JJ
中科院分区：
文献类型：
--
作者：
Lustberg MB;Balasubramanian P;Miller B;Garcia-Villa A;Deighan C;Wu Y;Carothers S;Berger M;Ramaswamy B;Macrae ER;Wesolowski R;Layman RM;Mrozek E;Pan X;Summers TA;Shapiro CL;Chalmers JJ
循环肿瘤细胞（CTCs）通常通过阳性选择靶向上皮细胞粘附分子（EpCAM）从血液中分离出来。然而，EpCAM在转移过程中可能下调，或者最初不存在。我们设计了目前的前瞻性试验，在没有epcam依赖性富集和/或分离技术的情况下，对转移性乳腺癌女性血液样本中的ctc以及其他循环细胞群进行表征。总共招募了32名转移性乳腺癌患者，血液样本使用先前描述的负耗竭免疫磁方法进行处理。健康志愿者的样本作为对照（n = 5）。进行多步骤顺序标记，标记和固定细胞表面标记物，然后渗透细胞角蛋白（CK） 8、18和19。采用BD LSR II型流式细胞仪或BD FACSAria II或FACSAria III型细胞分选仪进行多参数流式细胞仪（FCM）分析。对富集后的标本进行DAPI、EpCAM、CD45、CK、表皮生长因子受体和波形蛋白的免疫细胞化学染色。用共聚焦显微镜观察这些标记物的表达。CD45阴性/CK阳性（CD45−CK+）表达EpCAM +和EpCAM−的群体用FCM和CM从阴性富集的患者样本中鉴定。此外，还发现了CK +和共表达泛造血标志物CD45的EpCAM +和EpCAM−群体。CD45 -和CD45+组CK + EpCAM - events/ml均多于CK + EpCAM + events/ml （P≤0.0005均有统计学意义）。患者样本中每毫升血液样本（无论EpCAM状态如何）中CK + CD45−和CK + CD45+事件的数量高于正常对照样本（P分别≤0.0005和P≤0.026）。此外，CK + CD45+事件的很大一部分也表达CD68，这是一种与肿瘤相关巨噬细胞相关的标志物。较高水平的CD45-CK + EpCAM−与较差的总生存率相关（P = 0.0292）。转移性乳腺癌患者血液中存在CK + EpCAM−的非典型细胞。由于这些患者中有相当一部分没有EpCAM + ctc，因此需要进一步的研究来评估EpCAM -循环细胞作为预后和预测标志物的作用。
Circulating tumor cells (CTCs) are commonly isolated from the blood by targeting the epithelial cell adhesion molecule (EpCAM) through positive selection. However, EpCAM can be downregulated during metastatic progression, or it can be initially not present. We designed the present prospective trial to characterize CTCs as well as other circulating cell populations in blood samples from women with metastatic breast cancer without EpCAM-dependent enrichment and/or isolation technology. A total of 32 patients with metastatic breast cancer were enrolled, and blood samples were processed using a previously described negative depletion immunomagnetic methodology. Samples from healthy volunteers were run as controls (n = 5). Multistep sequential labeling was performed to label and fix cell-surface markers followed by permeabilization for cytokeratins (CK) 8, 18 and 19. Multiparametric flow cytometry (FCM) analysis was conducted using a BD LSR II flow cytometer or a BD FACSAria II or FACSAria III cell sorter. Immunocytochemical staining on postenrichment specimens for DAPI, EpCAM, CD45, CK, epidermal growth factor receptor and vimentin was performed. Expression of these markers was visualized using confocal microscopy (CM). CD45-negative/CK-positive (CD45− CK+) populations with EpCAM + and EpCAM − expression were identified with both FCM and CM from the negatively enriched patient samples. In addition, EpCAM + and EpCAM − populations that were CK + and coexpressing the pan-hematopoietic marker CD45 were also noted. There were more CK + EpCAM − events/ml than CK + EpCAM + events/ml in both the CD45− and CD45+ fractions (both statistically significant at P ≤ 0.0005). The number of CK + CD45− and CK + CD45+ events per milliliter in blood samples (regardless of EpCAM status) was higher in patient samples than in normal control samples (P ≤ 0.0005 and P ≤ 0.026, respectively). Further, a significant fraction of the CK + CD45+ events also expressed CD68, a marker associated with tumor-associated macrophages. Higher levels of CD45-CK + EpCAM − were associated with worse overall survival (P = 0.0292). Metastatic breast cancer patients have atypical cells that are CK + EpCAM − circulating in their blood. Because a substantial number of these patients do not have EpCAM + CTCs, additional studies are needed to evaluate the role of EpCAM − circulating cells as a prognostic and predictive marker.
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影响因子：
3.7
作者：
Balasubramanian P;Lang JC;Jatana KR;Miller B;Ozer E;Old M;Schuller DE;Agrawal A;Teknos TN;Summers TA Jr;Lustberg MB;Zborowski M;Chalmers JJ
通讯作者：
Chalmers JJ
影响因子：
158.5
作者：
Cristofanilli, M;Budd, GT;Hayes, DF
通讯作者：
Hayes, DF
影响因子：
11.5
作者：
Hayes, Daniel F.;Cristofanilli, Massimo;Terstappen, Leon W. W. M.
通讯作者：
Terstappen, Leon W. W. M.
影响因子：
--
作者：
Jatana, Kris R.;Balasubramanian, Priya;Lang, Jas C.;Yang, Liying;Jatana, Courtney A.;White, Elisabeth;Agrawal, Amit;Ozer, Enver;Schuller, David E.;Teknos, Theodoros N.;Chalmers, Jeffrey J.
通讯作者：
Chalmers, Jeffrey J.
影响因子：
50.5
作者：
Giordano, A.;Giuliano, M.;Cristofanilli, M.
通讯作者：
Cristofanilli, M.