Hendricks Alexander, Eggebrecht Greta-Lou, Bernsmeier Alexander, Geisen Reinhild, Dall Katharina, Trauzold Anna, Becker Thomas, Kalthoff Holger, Schafmayer Clemens, Röder Christian, Hinz Sebastian
Department of General and Thoracic Surgery, University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany.
Institute for Experimental Cancer Research, Christian-Albrechts University, Kiel, Germany.
Oncotarget. 2018 Jun 8;9(44):27423-27434. doi: 10.18632/oncotarget.25384.
In recent years, the concept of liquid biopsy diagnostics in detection and progress monitoring of malignant diseases gained significant awareness. We here report on a semi-quantitative real-time cytokeratin 20 RT-PCR-based assay, for detecting circulating tumor cells within a fraction of peripheral blood mononuclear cells in colorectal cancer patients.
In total, 381 patients were included. Prior to surgical tumor resection, a peripheral blood sample was drawn. Mononuclear cells were isolated by Ficoll centrifugation and a cytokeratin 20 qRT-PCR assay was performed. Quantitative PCR data was assessed regarding histopathological characteristics and patients´ clinical outcome.
A cut-off value was determined at ≥ 2.77 [EU]. Stratifying patients by this cut-off, it represents a statistically highly significant prognostic marker for both the overall and disease-free survival in the entire cohort UICC I-IV (both p<0.001) and in early tumor stages UICC I+II (overall survival p=0.003 and disease-free survival p=0.005). In multivariate analysis, the cut-off value stands for an independent predictor of significantly worse overall and disease-free survival (p=0.035 and p=0.047, respectively).
We successfully established a highly sensitive real-time qRT-PCR assay by which we are able to identify colorectal cancer patients at risk for an unfavorable prognosis in UICC I and II stages.
近年来，液体活检诊断在恶性疾病检测和进展监测中的概念得到了广泛关注。我们在此报告一种基于半定量实时细胞角蛋白20 RT-PCR的检测方法，用于检测结直肠癌患者外周血单个核细胞中的循环肿瘤细胞。
共纳入381例患者。在手术切除肿瘤前采集外周血样本。通过Ficoll离心分离单个核细胞，并进行细胞角蛋白20 qRT-PCR检测。评估定量PCR数据与组织病理学特征和患者临床结局的关系。
确定临界值为≥2.77 [EU]。按此临界值对患者进行分层，它是整个UICC I-IV队列（p均<0.001）以及早期肿瘤阶段UICC I+II（总生存p=0.003，无病生存p=0.005）中总生存和无病生存的统计学高度显著的预后标志物。在多变量分析中，该临界值是总生存和无病生存显著较差的独立预测因子（分别为p=0.035和p=0.047）。
我们成功建立了一种高度灵敏的实时qRT-PCR检测方法，通过该方法能够识别UICC I期和II期预后不良风险的结直肠癌患者。
