Chalfin Heather J, Kates Max, van der Toom Emma E, Glavaris Stephanie, Verdone James E, Hahn Noah M, Pienta Kenneth J, Bivalacqua Trinity J, Gorin Michael A
The James Buchanan Brady Urological Institute and Department of Urology, Johns Hopkins University School of Medicine, Baltimore, MD.
The James Buchanan Brady Urological Institute and Department of Urology, Johns Hopkins University School of Medicine, Baltimore, MD.
Urology. 2018 May;115:82-86. doi: 10.1016/j.urology.2018.01.036. Epub 2018 Feb 9.
To investigate circulating tumor cells (CTCs) as biomarkers of urothelial carcinoma (UC). To date, the majority of work on this topic has utilized the CellSearch test, which has limited sensitivity due to reliance on positive selection for the cell surface protein epithelial cell adhesion molecule (EpCAM). We used a novel selection-free method to enumerate and characterize CTCs across a range of UC stages.
Blood samples from 38 patients (9 controls, 8 nonmuscle invasive bladder cancer [NMIBC], 12 muscle-invasive bladder cancer [MIBC], and 9 metastatic UC) were processed with the AccuCyte-CyteFinder system. Slides were stained for the white blood cell markers CD45 and CD66b and the epithelial markers EpCAM and pancytokeratin. CTCs were defined as any cytokeratin postive and white blood cell marker negative cell. Separately, the more restrictive CellSearch definition was applied, with the additional requirement of EpCAM positivity. The Kruskal-Wallis ANOVA test compared CTC counts by stage.
Greater than or equal to 1 CTC was detected in 2 of 8 (25%) patients with NMIBC, 7 of 12 (58%) with MIBC, and 6of 9 (67%) with metastatic disease. No control had CTCs. Comparing CTC counts between groups, the only statistically significant comparison was between controls and patients with metastatic UC (P = .009). With EpCAM positivity as a CTC requirement, no CTCs were detected in any patient with NMIBC, and only 2 (17%) patients with MIBC had CTCs. CTCs tended to be larger in metastatic patients.
CTCs were detected at all UC stages and exhibited phenotypic diversity of cell size and EpCAM expression. EpCAM negative CTCs that would be missed with the CellSearch test were detected in patients with NMIBC and patients with MIBC.
研究循环肿瘤细胞（CTCs）作为尿路上皮癌（UC）生物标志物的情况。迄今为止，关于该主题的大多数研究都采用了CellSearch检测方法，由于依赖对细胞表面蛋白上皮细胞粘附分子（EpCAM）进行阳性选择，其灵敏度有限。我们使用一种新型的无选择方法来对一系列UC分期的CTCs进行计数和表征。
对38例患者（9例对照、8例非肌层浸润性膀胱癌[NMIBC]、12例肌层浸润性膀胱癌[MIBC]和9例转移性UC）的血样使用AccuCyte-CyteFinder系统进行处理。玻片用白细胞标志物CD45和CD66b以及上皮标志物EpCAM和全细胞角蛋白进行染色。CTCs定义为任何细胞角蛋白阳性且白细胞标志物阴性的细胞。另外，应用了更严格的CellSearch定义，即还需EpCAM呈阳性。采用Kruskal-Wallis方差分析检验按分期比较CTCs计数。
8例NMIBC患者中有2例（25%）检测到≥1个CTCs，12例MIBC患者中有7例（58%），9例转移性疾病患者中有6例（67%）。对照组未检测到CTCs。比较各组间的CTCs计数，唯一具有统计学意义的比较是对照组与转移性UC患者之间（P = 0.009）。以EpCAM阳性作为CTCs的标准时，任何NMIBC患者均未检测到CTCs，仅2例（17%）MIBC患者有CTCs。转移性患者的CTCs往往更大。
在所有UC分期均检测到CTCs，且其细胞大小和EpCAM表达呈现表型多样性。在NMIBC患者和MIBC患者中检测到了CellSearch检测会遗漏的EpCAM阴性CTCs。
