Abstract
Background
A growing body of research indicates that the monitoring of circulating tumor cells (CTCs) may have great significance to the diagnosis of malignant tumors, assessment of condition, selection of treatment methods, and evaluation of prognosis and has a broad range of potential applications. However, the value of CTCs with different phenotypes in the diagnosis of hepatocellular carcinoma (HCC) and assessment of patient condition remains unclear.
Methods
We collected 5 ml of peripheral blood from 176 patients who were found to have space-occupying lesions in the liver via B-ultrasound diagnosis at Zhujiang Hospital affiliated with Southern Medical University between August 2015 and October 2017 and used CanPatrol™ CTCs assay technology to isolate and count CTCs with different phenotypes in the patients’ peripheral blood. This allowed analysis of the value of CTCs with different phenotypes in the diagnosis of HCC and assessment of BCLC stage.
Results
We used CanPatrol™ CTCs assay technology to isolate different types of CTCs: epithelial CTCs (only stained for epithelial markers), mesenchymal CTCs (only stained for mesenchymal markers), mixed CTCs (stained for epithelial markers and mesenchymal markers), and total CTCs (all of the foregoing CTC phenotypes). Of 176 observed patients, 6 patients were finally diagnosed as other malignant tumor liver metastasis, 113 were diagnosed as having hepatocellular carcinoma, and 57 were diagnosed as having nonmalignant liver diseases. Furthermore, we intend to evaluate the diagnostic value of different phenotype CTCs count in discrimination between hepatocellular carcinoma and nonmalignant liver diseases. We found that CTCs of all types were significantly more numerous in the peripheral blood of the HCC group patients than in the NLD group patients (P < 0.05). Furthermore, of the different types of CTCs, total CTCs had the greatest diagnostic value (AUC 0.774; 95% CI, 0.704–0.834). A further discovery was that the AUC values for total CTCs, AFP, and a combined model (combined use of total CTCs and AFP) were 0.774 (95%CI, 0.704–0.834), 0.669 (95%CI, 0.587–0.750), and 0.821 (95%CI, 0.756–0.886). Late-stage HCC patients (BCLC stage B-C) had a higher peripheral blood mesenchymal CTC count than early-stage patients (BCLC stage 0-A) (median:1 vs 0), and mesenchymal CTCs ≥ 1 was the cut-off value for the diagnosis of BCLC stage in HCC patients (sensitivity: 66.67%, specificity: 59.46%, Youden index: 0.26).
Conclusions
Total CTCs are more effective than AFP in the diagnosis of HCC; combined use of total CTCs and AFP can enhance the sensitivity of HCC diagnosis.
Similar content being viewed by others
References
Ferenci P, Fried M, Labrecque D et al. World Gastroenterology Organisation Guideline. Hepatocellular carcinoma (HCC): a global perspective. J Gastrointestin Liver Dis 2010;19:311–317.
El-Serag H B, Rudolph K L. Hepatocellular carcinoma: epidemiology and molecular carcinogenesis. Gastroenterology 2007;132:2557–2576.
Yang B, Li M, Tang W et al. Dynamic network biomarker indicates pulmonary metastasis at the tipping point of hepatocellular carcinoma. Nat Commun 2018;9:678.
Erstad D J, Tanabe K K. Hepatocellular carcinoma: early-stage management challenges. J Hepatocell Carcinoma 2017;4:81–92.
Kudo M. Evidence and Consensus on the Management of Hepatocellular Carcinoma: Update 2015. Oncology 2015;89 Suppl 2:1–3.
Morimoto M, Numata K, Nozaki A et al. Novel Lens culinaris agglutinin-reactive fraction of α-fetoprotein: a biomarker of hepatocellular carcinoma recurrence in patients with low α-fetoprotein concentrations. International Journal of Clinical Oncology 2012;17:373–379.
Mehta N J, Celik A D, Peters M G. Screening for hepatocellular carcinoma: What is missing? Hepatology Communications 2017;1:18–22.
Zhang Y, Li J, Cao L et al. Circulating tumor cells in hepatocellular carcinoma: detection techniques, clinical implications, and future perspectives. Semin Oncol 2012;39:449–460.
Plaks V, Koopman C D, Werb Z. Circulating Tumor Cells. Science 2013;341:1186–1188.
Alix-Panabieres C, Pantel K. Circulating Tumor Cells: Liquid Biopsy of Cancer. Clinical Chemistry 2013;59:110–118.
Joosse S A, Gorges T M, Pantel K. Biology, detection, and clinical implications of circulating tumor cells. EMBO Mol Med 2015;7:1–11.
Pierga J Y, Bidard F C, Mathiot C et al. Circulating Tumor Cell Detection Predicts Early Metastatic Relapse After Neoadjuvant Chemotherapy in Large Operable and Locally Advanced Breast Cancer in a Phase II Randomized Trial. Clinical Cancer Research 2008;14:7004–7010.
Franken B, de Groot M R, Mastboom W J et al. Circulating tumor cells, disease recurrence and survival in newly diagnosed breast cancer. Breast Cancer Res 2012;14:R133.
Cristofanilli M. Circulating tumour cells: telling the truth about metastasis. Lancet Oncol 2014;15:365–366.
Liu Y, Hu B, Li Z et al. An improved strategy to detect the epithelial-mesenchymal transition process in circulating tumor cells in hepatocellular carcinoma patients. Hepatology International 2016;10:640–646.
Wang Z, Luo L, Cheng Y et al. Correlation Between Postoperative Early Recurrence of Hepatocellular Carcinoma and Mesenchymal Circulating Tumor Cells in Peripheral Blood. Journal of Gastrointestinal Surgery 2018;,22:633–639.
Kikuchi L, Oliveira C P, Alvares-Da-Silva M R et al. Hepatocellular Carcinoma Management in Nonalcoholic Fatty Liver Disease Patients: Applicability of the BCLC Staging System. Am J Clin Oncol 2016;39:428–432.
Li T. Evaluation of epithelial-mesenchymal transitioned circulating tumor cells in patients with resectable gastric cancer: Relevance to therapy response. World Journal of Gastroenterology 2015;21:13259.
Sun Y F, Xu Y, Yang X R et al. Circulating stem cell-like epithelial cell adhesion molecule-positive tumor cells indicate poor prognosis of hepatocellular carcinoma after curative resection. Hepatology 2013;57:1458–1468.
Riethdorf S, O'Flaherty L, Hille C et al. Clinical applications of the CellSearch platform in cancer patients. Adv Drug Deliv Rev 2018; 125:102–121
Barrière G, Tartary M, Rigaud M. Epithelial Mesenchymal Transition: A New Insight into the Detection of Circulating Tumor Cells. ISRN Oncology 2012;2012:1–6.
Giesing M, Driesel G, Molitor D et al. Molecular phenotyping of circulating tumour cells in patients with prostate cancer: prediction of distant metastases. BJU Int 2012;110:E1202-E1211.
Krawczyk N, Meier-Stiegen F, Banys M et al. Expression of stem cell and epithelial-mesenchymal transition markers in circulating tumor cells of breast cancer patients. Biomed Res Int 2014;2014:415721.
Gradilone A, Naso G, Raimondi C et al. Circulating tumor cells (CTCs) in metastatic breast cancer (MBC): prognosis, drug resistance and phenotypic characterization. Ann Oncol 2011;22:86–92.
Yu M, Bardia A, Wittner B S et al. Circulating Breast Tumor Cells Exhibit Dynamic Changes in Epithelial and Mesenchymal Composition. Science 2013;339:580–584.
Sun T, Zhao N, Zhao X L et al. Expression and functional significance of Twist1 in hepatocellular carcinoma: its role in vasculogenic mimicry. Hepatology 2010: 51: 545–556.
Sun Y, Guo W, Xu Y et al. Circulating Tumor Cells from Different Vascular Sites Exhibit Spatial Heterogeneity in Epithelial and Mesenchymal Composition and Distinct Clinical Significance in Hepatocellular Carcinoma. Clinical Cancer Research 2018;24:547–559.
Wu F, Zhu J, Mao Y et al. Associations between the Epithelial-Mesenchymal Transition Phenotypes of Circulating Tumor Cells and the Clinicopathological Features of Patients with Colorectal Cancer. Disease Markers 2017;2017:1–6.
Liu D, Xue L, Li J et al. Epithelial-mesenchymal transition and GALC expression of circulating tumor cells indicate metastasis and poor prognosis in non-small cell lung cancer. Cancer Biomarkers 2018;22:417–426.
Micalizzi D S, Maheswaran S, Haber D A. A conduit to metastasis: circulating tumor cell biology. Genes Dev 2017;31:1827–1840.
Raimondi C, Carpino G, Nicolazzo C et al. PD-L1 and epithelial-mesenchymal transition in circulating tumor cells from non-small cell lung cancer patients: A molecular shield to evade immune system? OncoImmunology 2017;6:e1315488.
Lianidou E S, Strati A, Markou A. Circulating tumor cells as promising novel biomarkers in solid cancers. Critical Reviews in Clinical Laboratory Sciences 2014; 51: 160–171.
Bai D S, Zhang C, Chen P et al. The prognostic correlation of AFP level at diagnosis with pathological grade, progression, and survival of patients with hepatocellular carcinoma. Sci Rep 2017;7:12870.
Fateen W, Ryder S D. Screening for hepatocellular carcinoma: patient selection and perspectives. J Hepatocell Carcinoma 2017;4:71–79.
Tanaka F, Yoneda K, Kondo N, et al. Circulating tumor cell as a diagnostic marker in primary lung cancer. Clin Cancer Res 2009;15:6980–6986.
Lin J, Zhang Y, Wu J et al. Neuropilin 1 (NRP1) is a novel tumor marker in hepatocellular carcinoma. Clin Chim Acta 2018;485:158–165.
Choi J, Kim G A, Han S et al. Longitudinal Assessment of Three Serum Biomarkers to Detect Very Early Stage Hepatocellular Carcinoma. Hepatology 2018. [Epub ahead of print].
Li Y, Xu S, Li J et al. Epithelial–mesenchymal transition markers expressed in circulating tumor cells in hepatocellular carcinoma patients with different stages of disease. Cell Death & Disease 2013;4:e831.
Akamatsu N, Cillo U, Cucchetti A et al. Surgery and Hepatocellular Carcinoma. Liver Cancer 2016;6:44–50.
Acknowledgments
We thank Surexam Biotech (Guangzhou, China) for the technical support. We are grateful to all the patients at Zhujiang Hospital who participated in this study.
Availability of Data and Materials
Please contact author for data requests.
Funding
This work was supported by grants from Natural Science Foundation of Guangdong Province, China, No.2016A030313626. The funding had no role in the collection, analysis, and interpretation of data, design of the study, or writing of the manuscript.
Author information
Authors and Affiliations
Contributions
Yuan Cheng, Lei Luo, and Juqiang Zhang contributed equally to this work and should be considered as co-first authors. Yuan Cheng, Lei Luo, Juqiang Zhang, and Mantian Zhou performed the experiments. Yujun Tang, Guolin He, and Yishi Lu collected patients’ information. Zhong Wang and MingXin Pan designed the study, edited the manuscript, and confirmed the data presented in the manuscript. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Competing Interests
The authors declare that they have no competing interests.
Ethics Approval and Consent to Participate
This study protocol was approved by the Institutional Review Board of the Second Affiliated Hospital of Southern Medical University (ZJYY-2015-GDEK-001). All participants will provide informed written consent prior to their entry into the study. In the case that changes in the protocol are necessary, relevant amendments will be made and submitted to the ethics trial registration authorities for approval.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Cheng, Y., Luo, L., Zhang, J. et al. Diagnostic Value of Different Phenotype Circulating Tumor Cells in Hepatocellular Carcinoma. J Gastrointest Surg 23, 2354–2361 (2019). https://doi.org/10.1007/s11605-018-04067-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11605-018-04067-y