Detection of circulating tumor cells in hepatocellular carcinoma using antibodies against asialoglycoprotein receptor, carbamoyl phosphate synthetase 1 and pan-cytokeratin.

Bibliographic Details
Title: Detection of circulating tumor cells in hepatocellular carcinoma using antibodies against asialoglycoprotein receptor, carbamoyl phosphate synthetase 1 and pan-cytokeratin.
Authors: Jun Li, Lei Chen, Xiaofeng Zhang, Yu Zhang, Huiying Liu, Bin Sun, Linlin Zhao, Naijian Ge, Haihua Qian, Yefa Yang, Mengchao Wu, Zhengfeng Yin
Source: PLoS ONE, Vol 9, Iss 4, p e96185 (2014)
Publisher Information: Public Library of Science (PLoS), 2014.
Publication Year: 2014
Collection: LCC:Medicine
LCC:Science
Subject Terms: Medicine, Science
More Details: BackgroundAsialoglycoprotein receptor (ASGPR)-ligand-based separation combined with identification with Hep Par 1 or pan-cytokeratin (P-CK) antibody have been demonstrated to detect circulating tumor cells (CTCs) in hepatocellular carcinoma (HCC). The aim of this study was to develop an improved enrichment and identification system that allows the detection of all types of HCC CTCs.MethodsThe specificity of the prepared anti-ASGPR monoclonal antibody was characterized. HCC cells were bound by ASGPR antibody and subsequently magnetically isolated by second antibody-coated magnetic beads. Isolated HCC cells were identified by immunofluorescence staining using a combination of anti-P-CK and anti-carbamoyl phosphate synthetase 1 (CPS1) antibodies. Blood samples spiked with HepG2 cells were used to determine recovery and sensitivity. CTCs were detected in blood samples from HCC patients and other patients.ResultsASGPR was exclusively expressed in human hepatoma cell line, normal hepatocytes and HCC cells in tissue specimens detected by the ASGPR antibody staining. More HCC cells could be identified by the antibody cocktail for CPS1 and P-CK compared with a single antibody. The current approach obtained a higher recovery rate of HepG2 cells and more CTC detection from HCC patients than the previous method. Using the current method CTCs were detected in 89% of HCC patients and no CTCs were found in the other test subjects.ConclusionsOur anti-ASGPR antibody could be used for specific and efficient HCC CTC enrichment, and anti-P-CK combined with anti-CPS1 antibodies is superior to identification with one antibody alone in the sensitivity for HCC CTC detection.
Document Type: article
File Description: electronic resource
Language: English
ISSN: 1932-6203
Relation: https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0096185&type=printable; https://doaj.org/toc/1932-6203
DOI: 10.1371/journal.pone.0096185&type=printable
DOI: 10.1371/journal.pone.0096185
Access URL: https://doaj.org/article/f47a210109be4aaa9c5618c003ff7ace
Accession Number: edsdoj.f47a210109be4aaa9c5618c003ff7ace
Database: Directory of Open Access Journals
More Details
ISSN:19326203
DOI:10.1371/journal.pone.0096185&type=printable
Published in:PLoS ONE
Language:English