Comparison of isolation platforms for detection of circulating renal cell carcinoma cells.
Comparison of isolation platforms for detection of circulating renal cell carcinoma cells.
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DOI：
10.18632/oncotarget.21197
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发表时间：
2017-10-20
期刊：
影响因子：
--
通讯作者：
Bernemann C
中科院分区：
文献类型：
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作者：
Maertens Y;Humberg V;Erlmeier F;Steffens S;Steinestel J;Bögemann M;Schrader AJ;Bernemann C
循环肿瘤细胞 (CTC) 的分析在多种肿瘤实体中取得了进展。然而，人们对透明细胞肾细胞癌 (ccRCC) 患者的 CTC 知之甚少。我们研究的目的是为 ccRCC 中 CTC 的分离建立稳定的体外基础。我们比较了不同 CTC 分离方法在产量和纯度方面的分析性能：基于 EpCAM 的富集、白细胞去除和基于大小的富集。在 ccRCC 细胞系和临床样本中评估了作为 CTC 表达生物标志物的 EpCAM 和细胞角蛋白 8 (KRT8)。虽然基于 EpCAM 的方法未能成功分离肿瘤细胞，但基于 CD45 的方法显示出中等的恢复率。基于细胞大小的 Parsortix 系统显示出最高的回收率。大多数细胞系以及临床样本中 EpCAM 表达较低或不表达，而 KRT8 被检测为 ccRCC 中的潜在生物标志物。基于 EpCAM 的方法可能会由于 ccRCC 中 EpCAM 表达低或缺失而错过大量 CTC，如细胞系和患者样本中所示。我们确定基于细胞大小、独立于标签的 Parsortix 系统是 ccRCC CTC 最有效的回收系统。
Analysis of circulating tumor cells (CTCs) has progressed in several tumor entities. However, little is known about CTCs in clear cell renal cell carcinoma (ccRCC) patients. Aim of our studies was to build a stable in vitro fundament for isolation of CTCs in ccRCC. We compared the analytical performance of different CTC isolation methods with regard to yield and purity: EpCAM based enrichment, leukocyte depletion and size based enrichment. EpCAM and cytokeratin 8 (KRT8) as biomarker for CTCs expression were evaluated in ccRCC cell lines as well as clinical samples. While the EpCAM based approach failed to successfully isolate tumor cells, CD45 based approaches showed intermediate recovery rates. The cell-size based Parsortix system showed highest recovery rates. EpCAM expression was low or absent in most cell lines as well as in clinical samples, whereas KRT8 was detected as a potential biomarker in ccRCC. EpCAM based approaches might miss a high number of CTCs due to low or absent expression of EpCAM in ccRCC, as shown in cell lines as well as in patient samples. We identified the cell-sized based, label independent Parsortix system to be the most effective recovery system for ccRCC CTCs.