采用依赖和不依赖上皮标志物的方法分析非小细胞肺癌患者的循环肿瘤细胞。
Analysis of circulating tumor cells in patients with non-small cell lung cancer using epithelial marker-dependent and -independent approaches.
机构信息
Clinical and Experimental Pharmacology Group, Paterson Institute for Cancer Research, University of Manchester, United Kingdom.
出版信息
J Thorac Oncol. 2012 Feb;7(2):306-15. doi: 10.1097/JTO.0b013e31823c5c16.
INTRODUCTION
Epithelial circulating tumor cells (CTCs) are detectable in patients with non-small cell lung cancer (NSCLC). However, epithelial to mesenchymal transition, a widely reported prerequisite for metastasis, may lead to an underestimation of CTC number. We compared directly an epithelial marker-dependent (CellSearch) and a marker-independent (isolation by size of epithelial tumor cells [ISET]) technology platform for the ability to identify CTCs. Molecular characteristics of CTCs were also explored.
METHODS
Paired peripheral blood samples were collected from 40 chemonäive, stages IIIA to IV NSCLC patients. CTCs were enumerated by Epithelial Cell Adhesion Molecule-based immunomagnetic capture (CellSearch, Veridex) and by filtration (ISET, RareCell Diagnostics). CTCs isolated by filtration were assessed by immunohistochemistry for epithelial marker expression (cytokeratins, Epithelial Cell Adhesion Molecule, epidermal growth factor receptor) and for proliferation status (Ki67).
RESULTS
CTCs were detected using ISET in 32 of 40 (80%) patients compared with 9 of 40 (23%) patients using CellSearch. A subpopulation of CTCs isolated by ISET did not express epithelial markers. Circulating tumor microemboli (CTM, clusters of ≥ 3 CTCs) were observed in 43% patients using ISET but were undetectable by CellSearch. Up to 62% of single CTCs were positive for the proliferation marker Ki67, whereas cells within CTM were nonproliferative.
CONCLUSIONS
Both technology platforms detected NSCLC CTCs. ISET detected higher numbers of CTCs including epithelial marker negative tumor cells. ISET also isolated CTM and permitted molecular characterization. Combined with our previous CellSearch data confirming CTC number as an independent prognostic biomarker for NSCLC, we propose that this complementary dual technology approach to CTC analysis allows more complete exploration of CTCs in patients with NSCLC.
简介
上皮循环肿瘤细胞（CTCs）可在非小细胞肺癌（NSCLC）患者中检测到。然而，上皮-间充质转化是广泛报道的转移前提条件，可能导致对 CTC 数量的低估。我们直接比较了依赖上皮标志物（CellSearch）和非标志物依赖（上皮肿瘤细胞大小分离[ISET]）的技术平台识别 CTC 的能力。还探索了 CTC 的分子特征。
方法
采集 40 例未经化疗的 IIIA 至 IV 期 NSCLC 患者的配对外周血样本。通过基于上皮细胞黏附分子的免疫磁捕获（CellSearch，Veridex）和过滤（ISET，RareCell Diagnostics）计数 CTCs。通过免疫组织化学评估过滤分离的 CTCs 的上皮标志物表达（细胞角蛋白、上皮细胞黏附分子、表皮生长因子受体）和增殖状态（Ki67）。
结果
与 CellSearch 相比，ISET 检测到 40 例患者中的 32 例（80%）有 CTCs，而 CellSearch 检测到 9 例（23%）。ISET 分离的 CTC 亚群不表达上皮标志物。使用 ISET 观察到 43%的患者存在循环肿瘤微栓（CTC 簇≥3 个），而 CellSearch 无法检测到。高达 62%的单个 CTCs 增殖标志物 Ki67 阳性，而 CTCM 中的细胞则无增殖活性。
结论
两种技术平台均检测到 NSCLC CTCs。ISET 检测到更多数量的 CTCs，包括上皮标志物阴性的肿瘤细胞。ISET 还分离了 CTCM，并允许进行分子特征分析。结合我们之前使用 CellSearch 数据证实 CTC 数量是 NSCLC 的独立预后生物标志物，我们建议这种互补的 CTC 分析双重技术方法可更全面地探索 NSCLC 患者的 CTC。
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