Clinical Significance of EpCAM mRNA-Positive Circulating Tumor Cells in Hepatocellular Carcinoma by an Optimized Negative Enrichment and qRT-PCR–Based Platform

• Published in: Clinical cancer research Vol. 20; no. 18; pp. 4794 - 4805
• Main Authors: Guo, Wei, Yang, Xin-Rong, Sun, Yun-Fan, Shen, Min-Na, Ma, Xiao-Lu, Wu, Jiong, Zhang, Chun-Yan, Zhou, Yan, Xu, Yang, Hu, Bo, Zhang, Xin, Zhou, Jian, Fan, Jia
• Format: Journal Article
• Language: English
• Published: United States 15.09.2014
• Subjects: Antigens, Neoplasm - metabolism; Biomarkers, Tumor - analysis; Biomarkers, Tumor - metabolism; Carcinoma, Hepatocellular - blood; Carcinoma, Hepatocellular - mortality; Carcinoma, Hepatocellular - pathology; Cell Adhesion Molecules - metabolism; Epithelial Cell Adhesion Molecule; Female; Humans; Kaplan-Meier Estimate; Liver Neoplasms - blood; Liver Neoplasms - mortality; Liver Neoplasms - pathology; Male; Middle Aged; Neoplastic Cells, Circulating - metabolism; Neoplastic Cells, Circulating - pathology; Real-Time Polymerase Chain Reaction - methods; RNA, Messenger; Sensitivity and Specificity
• ISSN: 1078-0432; 1557-3265; 1557-3265
• DOI: 10.1158/1078-0432.CCR-14-0251

Purpose: This study aimed to construct a novel platform for the detection of circulating tumor cells (CTC) in patients with hepatocellular carcinoma (HCC) and to investigate the clinical significance of epithelial cell adhesion molecule mRNA-positive (EpCAMmRNA+) CTCs using this platform. Experimental Design: An optimized platform for CTC detection was constructed by evaluating different negative enrichment, mRNA isolation, and cDNA synthesis procedures and compared with the CellSearch system. A total of 299 patients with HCC were recruited into this prospective study; of these, 157 who received curative resection, 76 who received transcatheter arterial chemoembolization (TACE), and 66 who received radiotherapy were tested using our platform. The diagnostic value of EpCAMmRNA+ CTCs was investigated in 122 patients with HCC who underwent resection and 120 control subjects. Results: The optimized negative enrichment and quantitative real-time PCR (qRT-PCR)-based CTC detection platform had high sensitivity, specificity, and reproducibility and a low sample volume requirement. This platform showed a potential diagnostic value in patients with HCC and exhibited 76.7% consistency with the CellSearch system (r = 0.54, P < 0.050). Pretreatment CTC level showed prognostic significance in patients with HCC treated with resection, TACE, and radiotherapy (all P < 0.050). Most of the patients showed a decrease in CTC levels after treatment that reflected tumor response. In contrast, patients with an increased CTC level showed disease progression after treatment. Conclusions: We established an optimized platform based on negative enrichment and qRT-PCR for highly sensitive, specific, and reproducible CTC detection. This platform might be clinically useful in auxiliary diagnosis, treatment response assessment, and early decision-making to tailor the most effective antitumor strategies. Clin Cancer Res; 20(18); 4794–805. ©2014 AACR.