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

TitleEstablishment and characterization of a novel primary hepatocellular carcinoma cell line with metastatic ability in vivo
Authors
Issue Date2014
Citation
Cancer Cell International, 2014, v. 14 n. 1, p. 103 How to Cite?
AbstractBACKGROUND: 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.
Persistent Identifierhttp://hdl.handle.net/10722/207312
ISSN
2015 Impact Factor: 2.884
2015 SCImago Journal Rankings: 1.050
PubMed Central ID

 

DC FieldValueLanguage
dc.contributor.authorCheung, PFYen_US
dc.contributor.authorYip, CWen_US
dc.contributor.authorNg, WCLen_US
dc.contributor.authorLo, KWen_US
dc.contributor.authorWong, Nen_US
dc.contributor.authorChoy, KWen_US
dc.contributor.authorChow, Cen_US
dc.contributor.authorChan, KFen_US
dc.contributor.authorCheung, TTen_US
dc.contributor.authorPoon, RTPen_US
dc.contributor.authorFan, STen_US
dc.contributor.authorCheung, STen_US
dc.date.accessioned2014-12-19T10:20:49Z-
dc.date.available2014-12-19T10:20:49Z-
dc.date.issued2014en_US
dc.identifier.citationCancer Cell International, 2014, v. 14 n. 1, p. 103en_US
dc.identifier.issn1475-2867-
dc.identifier.urihttp://hdl.handle.net/10722/207312-
dc.description.abstractBACKGROUND: 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.-
dc.languageengen_US
dc.relation.ispartofCancer Cell Internationalen_US
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.titleEstablishment and characterization of a novel primary hepatocellular carcinoma cell line with metastatic ability in vivoen_US
dc.typeArticleen_US
dc.identifier.emailCheung, PFY: cphyllis@hkucc.hku.hken_US
dc.identifier.emailYip, CW: wallacey@hku.hken_US
dc.identifier.emailNg, WCL: lindanwc@hku.hken_US
dc.identifier.emailCheung, TT: cheung68@hku.hken_US
dc.identifier.emailPoon, RTP: poontp@hku.hken_US
dc.identifier.emailFan, ST: stfan@hku.hken_US
dc.identifier.emailCheung, ST: stcheung@hkucc.hku.hken_US
dc.identifier.authorityPoon, RTP=rp00446en_US
dc.identifier.authorityFan, ST=rp00355en_US
dc.identifier.authorityCheung, ST=rp00457en_US
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.1186/s12935-014-0103-yen_US
dc.identifier.pmid25349534-
dc.identifier.pmcidPMC4209051-
dc.identifier.hkuros241966en_US
dc.identifier.volume14en_US
dc.identifier.issue1en_US
dc.identifier.spage103en_US
dc.identifier.epage103en_US

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