Department of Experimental Physiology, Medical School, National and Kapodistrian University of Athens, Micras Asias 75, Goudi-Athens, 11527 Athens, Greece.
World J Gastroenterol. 2010 Dec 21;16(47):5965-74. doi: 10.3748/wjg.v16.i47.5965.
To develop a multiplex reverse transcription polymerase chain reaction (RT-PCR) method detecting circulating tumor cells in the peripheral blood of colorectal cancer (CRC) patients.
Peripheral blood samples were collected from 88 CRC patients and 40 healthy individuals from the blood donors' clinic and subsequently analyzed by multiplex RT-RCR for the expression of carcinoembryonic antigen (CEA), cytokeratin 20 (CK20) and epidermal growth factor receptor (EGFR) mRNA. The analysis involved determining the detection rates of CEA, CK20 and EGFR transcripts vs disease stage and overall survival. Median follow-up period was 19 mo (range 8-28 mo).
Rates of CEA, CK20 and EGFR detection in CRC patients were 95.5%, 78.4% and 19.3%, respectively. CEA transcripts were detected in 3 healthy volunteer samples (7.5%), whereas all control samples were tested negative for CK20 and EGFR transcripts. The increasing number of positive detections for CEA, CK20 and EGFR transcripts in each blood sample was positively correlated with Astler-Coller disease stage (P < 0.001) and preoperative serum levels of CEA (P = 0.029) in CRC patients. Data analysis using Kaplan-Meier estimator documented significant differences in the overall survival of the different CRC patient groups as formed according to the increasing number of positivity for CEA, CK20 and EGFR transcripts.
These data suggest that multiplex RT-PCR assay can provide useful information concerning disease stage and overall survival of CRC patients.
开发一种用于检测结直肠癌（CRC）患者外周血循环肿瘤细胞的多重逆转录聚合酶链反应（RT-PCR）方法。
收集 88 例 CRC 患者和 40 例健康献血者的外周血样本，随后通过多重 RT-PCR 检测癌胚抗原（CEA）、细胞角蛋白 20（CK20）和表皮生长因子受体（EGFR）mRNA 的表达。分析包括检测 CEA、CK20 和 EGFR 转录本与疾病分期和总生存期的关系。中位随访时间为 19 个月（范围 8-28 个月）。
CRC 患者 CEA、CK20 和 EGFR 的检测率分别为 95.5%、78.4%和 19.3%。CEA 转录本在 3 例健康志愿者样本中检测到（7.5%），而所有对照样本均检测到 CK20 和 EGFR 转录本阴性。每个血液样本中 CEA、CK20 和 EGFR 转录本的阳性检出数与 Astler-Coller 疾病分期（P < 0.001）和术前 CEA 血清水平（P = 0.029）呈正相关。Kaplan-Meier 估计器数据分析显示，根据 CEA、CK20 和 EGFR 转录本阳性检出数的增加，CRC 患者的总生存期存在显著差异。
这些数据表明，多重 RT-PCR 检测方法可以为 CRC 患者的疾病分期和总生存期提供有用的信息。
