Circulating stem cell–like epithelial cell adhesion molecule–positive tumor cells indicate poor prognosis of hepatocellular carcinoma after curative resection

Abstract

Epithelial cell adhesion molecule-positive (EpCAM 1 ) hepatocellular carcinoma (HCC) cells may constitute a tumor-initiating subpopulation in tumorigenic cell lines and HCC specimens. In the present study, EpCAM 1 circulating tumor cells (CTCs) were identified prospectively in HCC patients undergoing curative resection, and the prognostic significance and their stem cell-like characteristics were investigated further. Blood samples from 123 HCC patients were tested prior to resection and 1 month thereafter. CTCs were present in 66.67% of patients, and the cell count measured in 7.5 mL of blood (CTC 7.5 ) ranged between 1 and 34. Fifty-one patients had CTC 7.5 of !2 preoperatively, and these patients developed tumor recurrence earlier than those with CTC 7.5 of <2 CTCs (P < 0.001). A preoperative CTC 7.5 of !2 was an independent prognostic factor for tumor recurrence (P < 0.001). Its prognostic significance also applied to patients with alpha-fetoprotein (AFP) levels of 400 ng/mL or subgroups with low recurrence risk (all P < 0.05). A significant decrease of CTC-positive rates (66.67% to 28.15%, P < 0.05) and CTC 7.5 values (2.60 6 0.43 to 1.00 6 0.36, P < 0.05) was observed 1 month after resection. Patients with consistent CTC 7.5 <2 had lower recurrence rates than those with values consistently !2 (15.5% versus 87.50%, P < 0.001). EpCAM 1 CTCs displayed cancer stem cell biomarkers (CD133 and ABCG2), epithelial-mesenchymal transition, Wnt pathway activation, high tumorigenic potential, and low apoptotic propensity. Conclusion: Stem cell-like phenotypes are observed in EpCAM 1 CTCs, and a preoperative CTC 7.5 of !2 is a novel predictor for tumor recurrence in HCC patients after surgery, especially in patient subgroups with AFP levels of 400 ng/mL or low tumor recurrence risk. EpCAM 1 CTCs may serve as a real-time parameter for monitoring treatment response and a therapeutic target in HCC recurrence. (HEPATOLOGY 2013;57:1458-1468 H epatocellular carcinoma (HCC) is one of the most prevalent malignancies worldwide, and associated morbidity and mortality rates have escalated in recent years. 1 Despite improvements in surveillance and clinical treatment strategies, the prognosis of HCC remains very poor due to high incidence of recurrence and metastasis. 2 Traditional clinicopathological parameters such as tumor morphology,

Detection of circulating tumor cells in pulmonary venous blood for resectable non‑small cell lung cancer

“…In the present study, the results of a xenograft assay demonstrated that the CTCs and CTM detected in the PV blood of patients were tumorigenic. Similar human CTC-derived xenograft models have been reported for breast cancer, hepatocellular carcinoma and SCLC (25)(26)(27). In addition, CTC culture has been established successfully from patients with early-stage NSCLC (28).…”

Circulating Tumor Cells from Different Vascular Sites Exhibit Spatial Heterogeneity in Epithelial and Mesenchymal Composition and Distinct Clinical Significance in Hepatocellular Carcinoma

“…The entrance criteria were: (i) definitive pathologic or radiologic diagnosis according to American Association for Study of Liver Disease guidelines; (ii) no extrahepatic metastasis at the time of diagnosis; (iii) no prior anticancer treatment (14). Tumor stage was determined according the Barcelona Clinic Liver Cancer (BCLC) staging system, and tumor differentiation was defined according to the Edmondson grading system.…”

“…The criteria for an EpCAM-positive object to be identified as a CTC were nucleated (DAPI þ ) intact cell, positive for pan-cytokeratin 8,18,19, and negative for CD45 (14). To prevent false assignment of a mitotic CTC as a microembolus, circulating tumor microemboli (CTM) was defined as clusters of CTCs containing three or more distinct nuclei (19).…”

“…We previously reported that peripheral venous CTCs detected by CELLSEARCH in HCC patients displayed considerable variation in epithelial-mesenchymal transition (EMT) features (14). In this study, we applied a single-cell RNA expression assay to further profile EMT-associated genes in individual CTCs captured by CELLSEARCH system.…”

“…In this study, CTCs were identified and characterized in gross systemic and hepatic vascular compartments as well as microscopic hepatic vasculatures along the blood flow pathway. The CELLSEARCH assay, considered the gold standard in CTC enumeration and confirmed by our previous studies (12)(13)(14), was used to investigate the differential localization of CTCs along their spreading route. The biological characteristics and potential clinical relevance of CTCs in different vascular compartments was further investigated, which was not reported previously.…”