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Cancer Cell International
Published in: Cancer Cell International 1/2014

Open Access 01-12-2014 | Primary research

Establishment and characterization of a novel primary hepatocellular carcinoma cell line with metastatic ability in vivo

Authors: Phyllis Fung-Yi Cheung, Chi Wai Yip, Linda Wing-Chi Ng, Kwok Wai Lo, Nathalie Wong, Kwong Wai Choy, Chit Chow, Kui Fat Chan, Tan To Cheung, Ronnie Tung-Ping Poon, Sheung Tat Fan, Siu Tim Cheung

Published in: Cancer Cell International | Issue 1/2014

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Abstract

Background

Hepatocellular carcinoma (HCC) is a highly aggressive and heterogeneous disease. HCC cell lines established from different patients would be useful in elucidating the molecular pathogenesis. However, success of HCC primary culture establishment remains at low rate. We aim to establish and characterize HCC primary culture and the derived cell line.

Methods

Fresh tumor tissues were collected from 30 HCC patients. Culture conditions were optimized for the attachment and growth of the isolated hepatocytes. Granulin-epithelin precursor (GEP), a growth factor reported to associate with cancer stem cell properties, was examined by flow cytometry to elucidate its role on primary culture establishment. The primary cell line was characterized in detail.

Results

Cells isolated from 16 out of 30 HCC cases (53%) had viability more than 70% and were subject to subsequent in vitro culture. 7 out of 16 cases (44%) could give rise to cells that were able to attach and grow in culture. GEP expression levels significantly correlated with the viability of isolated hepatocytes and success rate of subsequent primary culture establishment. Cells from HCC patient 21 grew and expanded rapidly in vitro and was selected to be further characterized. The line, designated HCC21, derived from a Hong Kong Chinese female patient with HCC at Stage II. The cells exhibited typical epithelial morphology and expressed albumin, AFP and HBV antigens. The cell line was authenticated by short tandem repeat analysis. Comparative genome hybridization analysis revealed chromosomal loss at 1p35-p36, 1q44, 2q11.2-q24.3, 2q37, 4q12-q13.3, 4q21.21-q35.2, 8p12-p23, 15q11.2-q14, 15q24-q26, 16p12.1-p13.3, 16q, 17p, 22q and gain at 1q21-q43 in both HCC21 cells and the original clinical tumor specimen. Sequence analysis revealed p53 gene mutation. Subcutaneous injection of HCC21 cells into immunodeficient mice showed that the cells were able to form tumors at the primary injection sites and metastatic tumors in the peritoneal cavity.

Conclusions

The newly established cell line could serve as useful in vitro and in vivo models for studying primary HCC that possess metastasis ability.
Appendix
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Metadata
Title
Establishment and characterization of a novel primary hepatocellular carcinoma cell line with metastatic ability in vivo
Authors
Phyllis Fung-Yi Cheung
Chi Wai Yip
Linda Wing-Chi Ng
Kwok Wai Lo
Nathalie Wong
Kwong Wai Choy
Chit Chow
Kui Fat Chan
Tan To Cheung
Ronnie Tung-Ping Poon
Sheung Tat Fan
Siu Tim Cheung
Publication date
01-12-2014
Publisher
BioMed Central
Published in
Cancer Cell International / Issue 1/2014
Electronic ISSN: 1475-2867
DOI
https://doi.org/10.1186/s12935-014-0103-y

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