Multiparameter analysis, including EMT markers, on negatively enriched blood samples from patients with squamous cell carcinoma of the head and neck.
Multiparameter analysis, including EMT markers, on negatively enriched blood samples from patients with squamous cell carcinoma of the head and neck.
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DOI：
10.1371/journal.pone.0042048
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Chalmers JJ
中科院分区：
文献类型：
--
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
上皮细胞向间质细胞转化（EMT）已被假设为细胞在癌变过程中改变表型以及肿瘤转移的机制。EMT是否参与癌症转移对循环肿瘤细胞（CTC）领域具有特定的实际影响。由于CTC的普遍接受的定义包括上皮表面标志物（例如EpCAM）的表达，如果癌细胞失去其上皮表面标志物（这在EMT中提出），则其将不会被分离和/或鉴定为CTC。我们已经开发了一种纯阴性富集技术，用于富集头颈部鳞状细胞癌（SCCHN）血液中的CTC，并且之前曾报道过该技术的使用。该方法不依赖于表面上皮标志物的表达。使用这项技术，我们对SCCHN患者血液的初步数据表明，CTC的存在与更差的无病生存率相关。由于我们的富集不依赖于上皮标志物，我们已经开始调查这些CTC细胞中存在的间充质标志物，包括分析：波形蛋白，表皮生长因子受体，N-钙粘蛋白和CD 44。借助共聚焦显微镜，我们不仅证明了假定的CTC表达和/或包含：细胞核，细胞角蛋白，波形蛋白，EGFR，CD 44或N-钙粘蛋白，而且还包含除细胞角蛋白外的所有上述蛋白质的细胞，这表明细胞经历了EMT过程。我们认为，我们的阴性耗竭富集方法提供了一种更客观的方法来鉴定和评估CTC，而不是阳性选择方法，因为它不受选择偏倚的影响，并且可以进行调整以适应各种细胞质和表面标志物，可以对这些标志物进行评价以鉴定来自个体患者的CTC内的多种表型模式，包括本文所述的所谓EMT。
Epithelial to mesenchymal transition (EMT) has been hypothesized as a mechanism by which cells change phenotype during carcinogenesis, as well as tumor metastasis. Whether EMT is involved in cancer metastasis has a specific, practical impact on the field of circulating tumor cells (CTCs). Since the generally accepted definition of a CTC includes the expression of epithelial surface markers, such as EpCAM, if a cancer cell loses its epithelial surface markers (which is suggested in EMT), it will not be separated and/or identified as a CTC. We have developed, and previously reported on the use of, a purely negative enrichment technology enriching for CTCs in the blood of squamous cell carcinoma of the head and neck (SCCHN). This methodology does not depend on the expression of surface epithelial markers. Using this technology, our initial data on SCCHN patient blood indicates that the presence of CTCs correlates with worse disease-free survival. Since our enrichment is not dependent on epithelial markers, we have initiated investigation of the presence of mesenchymal markers in these CTC cells to include analysis of: vimentin, epidermal growth factor receptor, N-cadherin, and CD44. With the aid of confocal microscopy, we have demonstrated not only presumed CTCs that express and/or contain: a nucleus, cytokeratins, vimentin, and either EGFR, CD44, or N-cadherin, but also cells that contain all of the aforementioned proteins except cytokeratins, suggesting that the cells have undergone the EMT process. We suggest that our negative depletion enrichment methodology provides a more objective approach in identifying and evaluating CTCs, as opposed to positive selection approaches, as it is not subjective to a selection bias and can be tailored to accommodate a variety of cytoplasmic and surface markers which can be evaluated to identify a multitude of phenotypic patterns within CTCs from individual patients, including so-called EMT as presented here.
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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：
2.7
作者：
Chaffer, Christine L.;Thompson, Erik W.;Williams, Elizabeth D.
通讯作者：
Williams, Elizabeth D.
影响因子：
11.2
作者：
Sarrió, David;Rodriguez-Pinilla, Socorro Maria;Palacios, José
通讯作者：
Palacios, José
影响因子：
4.9
作者：
Balasubramanian, Priya;Yang, Liying;Chalmers, Jeffrey J.
通讯作者：
Chalmers, Jeffrey J.
影响因子：
3.8
作者：
Yang, Liying;Lang, James C.;Balasubramanian, Priya;Jatana, Kris R.;Schuller, David;Agrawal, Amit;Zborowski, Maciej;Chalmers, Jeffrey J.
通讯作者：
Chalmers, Jeffrey J.