A long term, non-tumorigenic rat hepatocyte cell line and its malignant counterpart, as tools to study hepatocarcinogenesis

Maria Maddalena Angioni, Kevin Bellofatto, Simone Merlin, Silvia Menegon, Andrea Perra, Annalisa Petrelli, Pia Sulas, Silvia Giordano, Amedeo Columbano, Antonia Follenzi

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide and the second cause of cancer-related death. Search for genes/proteins whose expression can discriminate between normal and neoplastic liver is fundamental for diagnostic, prognostic and therapeutic purposes. Currently, the most used in vitro hepatocyte models to study molecular alterations underlying transformation include primary hepatocytes and transformed cell lines. However, each of these models presents limitations. Here we describe the isolation and characterization of two rat hepatocyte cell lines as tools to study liver carcinogenesis. Long-term stable cell lines were obtained from a HCC-bearing rat exposed to the Resistant-Hepatocyte protocol (RH cells) and from a rat subjected to the same model in the absence of carcinogenic treatment, thus not developing HCCs (RNT cells). The presence of several markers identified the hepatocytic origin of both cell lines and confirmed their purity. Although morphologically similar to normal primary hepatocytes, RNT cells were able to survive and grow in monolayer culture for months and were not tumorigenic in vivo. On the contrary, RH cells displayed tumor-initiating cell markers, formed numerous colonies in soft agar and spheroids when grown in 3D and were highly tumorigenic and metastatic after injection into syngeneic rats and immunocompromised mice. Moreover, RNT gene expression profile was similar to normal liver, while that of RH resembled HCC. In conclusion, the two cell lines here described represent a useful tool to investigate the molecular changes underlying hepatocyte transformation and to experimentally demonstrate their role in HCC development.

Original languageEnglish
Pages (from-to)15716-15731
Number of pages16
JournalOncotarget
Volume8
Issue number9
DOIs
Publication statusPublished - Jan 1 2017

Fingerprint

Hepatocytes
Cell Line
Hepatocellular Carcinoma
Liver
Transformed Cell Line
Second Primary Neoplasms
Neoplastic Stem Cells
Transcriptome
Agar
Carcinogenesis
Injections
Therapeutics
Neoplasms
Proteins

Keywords

  • CD24
  • CD90.1
  • HCC cell lines
  • Immortalized non-tumorigenic cells
  • Resistant hepatocyte

ASJC Scopus subject areas

  • Oncology

Cite this

Angioni, M. M., Bellofatto, K., Merlin, S., Menegon, S., Perra, A., Petrelli, A., ... Follenzi, A. (2017). A long term, non-tumorigenic rat hepatocyte cell line and its malignant counterpart, as tools to study hepatocarcinogenesis. Oncotarget, 8(9), 15716-15731. https://doi.org/10.18632/oncotarget.14984

A long term, non-tumorigenic rat hepatocyte cell line and its malignant counterpart, as tools to study hepatocarcinogenesis. / Angioni, Maria Maddalena; Bellofatto, Kevin; Merlin, Simone; Menegon, Silvia; Perra, Andrea; Petrelli, Annalisa; Sulas, Pia; Giordano, Silvia; Columbano, Amedeo; Follenzi, Antonia.

In: Oncotarget, Vol. 8, No. 9, 01.01.2017, p. 15716-15731.

Research output: Contribution to journalArticle

Angioni, MM, Bellofatto, K, Merlin, S, Menegon, S, Perra, A, Petrelli, A, Sulas, P, Giordano, S, Columbano, A & Follenzi, A 2017, 'A long term, non-tumorigenic rat hepatocyte cell line and its malignant counterpart, as tools to study hepatocarcinogenesis', Oncotarget, vol. 8, no. 9, pp. 15716-15731. https://doi.org/10.18632/oncotarget.14984
Angioni, Maria Maddalena ; Bellofatto, Kevin ; Merlin, Simone ; Menegon, Silvia ; Perra, Andrea ; Petrelli, Annalisa ; Sulas, Pia ; Giordano, Silvia ; Columbano, Amedeo ; Follenzi, Antonia. / A long term, non-tumorigenic rat hepatocyte cell line and its malignant counterpart, as tools to study hepatocarcinogenesis. In: Oncotarget. 2017 ; Vol. 8, No. 9. pp. 15716-15731.
@article{740fc1d01ec44dc0bcf920cb828348c2,
title = "A long term, non-tumorigenic rat hepatocyte cell line and its malignant counterpart, as tools to study hepatocarcinogenesis",
abstract = "Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide and the second cause of cancer-related death. Search for genes/proteins whose expression can discriminate between normal and neoplastic liver is fundamental for diagnostic, prognostic and therapeutic purposes. Currently, the most used in vitro hepatocyte models to study molecular alterations underlying transformation include primary hepatocytes and transformed cell lines. However, each of these models presents limitations. Here we describe the isolation and characterization of two rat hepatocyte cell lines as tools to study liver carcinogenesis. Long-term stable cell lines were obtained from a HCC-bearing rat exposed to the Resistant-Hepatocyte protocol (RH cells) and from a rat subjected to the same model in the absence of carcinogenic treatment, thus not developing HCCs (RNT cells). The presence of several markers identified the hepatocytic origin of both cell lines and confirmed their purity. Although morphologically similar to normal primary hepatocytes, RNT cells were able to survive and grow in monolayer culture for months and were not tumorigenic in vivo. On the contrary, RH cells displayed tumor-initiating cell markers, formed numerous colonies in soft agar and spheroids when grown in 3D and were highly tumorigenic and metastatic after injection into syngeneic rats and immunocompromised mice. Moreover, RNT gene expression profile was similar to normal liver, while that of RH resembled HCC. In conclusion, the two cell lines here described represent a useful tool to investigate the molecular changes underlying hepatocyte transformation and to experimentally demonstrate their role in HCC development.",
keywords = "CD24, CD90.1, HCC cell lines, Immortalized non-tumorigenic cells, Resistant hepatocyte",
author = "Angioni, {Maria Maddalena} and Kevin Bellofatto and Simone Merlin and Silvia Menegon and Andrea Perra and Annalisa Petrelli and Pia Sulas and Silvia Giordano and Amedeo Columbano and Antonia Follenzi",
year = "2017",
month = "1",
day = "1",
doi = "10.18632/oncotarget.14984",
language = "English",
volume = "8",
pages = "15716--15731",
journal = "Oncotarget",
issn = "1949-2553",
publisher = "Impact Journals LLC",
number = "9",

}

TY - JOUR

T1 - A long term, non-tumorigenic rat hepatocyte cell line and its malignant counterpart, as tools to study hepatocarcinogenesis

AU - Angioni, Maria Maddalena

AU - Bellofatto, Kevin

AU - Merlin, Simone

AU - Menegon, Silvia

AU - Perra, Andrea

AU - Petrelli, Annalisa

AU - Sulas, Pia

AU - Giordano, Silvia

AU - Columbano, Amedeo

AU - Follenzi, Antonia

PY - 2017/1/1

Y1 - 2017/1/1

N2 - Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide and the second cause of cancer-related death. Search for genes/proteins whose expression can discriminate between normal and neoplastic liver is fundamental for diagnostic, prognostic and therapeutic purposes. Currently, the most used in vitro hepatocyte models to study molecular alterations underlying transformation include primary hepatocytes and transformed cell lines. However, each of these models presents limitations. Here we describe the isolation and characterization of two rat hepatocyte cell lines as tools to study liver carcinogenesis. Long-term stable cell lines were obtained from a HCC-bearing rat exposed to the Resistant-Hepatocyte protocol (RH cells) and from a rat subjected to the same model in the absence of carcinogenic treatment, thus not developing HCCs (RNT cells). The presence of several markers identified the hepatocytic origin of both cell lines and confirmed their purity. Although morphologically similar to normal primary hepatocytes, RNT cells were able to survive and grow in monolayer culture for months and were not tumorigenic in vivo. On the contrary, RH cells displayed tumor-initiating cell markers, formed numerous colonies in soft agar and spheroids when grown in 3D and were highly tumorigenic and metastatic after injection into syngeneic rats and immunocompromised mice. Moreover, RNT gene expression profile was similar to normal liver, while that of RH resembled HCC. In conclusion, the two cell lines here described represent a useful tool to investigate the molecular changes underlying hepatocyte transformation and to experimentally demonstrate their role in HCC development.

AB - Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide and the second cause of cancer-related death. Search for genes/proteins whose expression can discriminate between normal and neoplastic liver is fundamental for diagnostic, prognostic and therapeutic purposes. Currently, the most used in vitro hepatocyte models to study molecular alterations underlying transformation include primary hepatocytes and transformed cell lines. However, each of these models presents limitations. Here we describe the isolation and characterization of two rat hepatocyte cell lines as tools to study liver carcinogenesis. Long-term stable cell lines were obtained from a HCC-bearing rat exposed to the Resistant-Hepatocyte protocol (RH cells) and from a rat subjected to the same model in the absence of carcinogenic treatment, thus not developing HCCs (RNT cells). The presence of several markers identified the hepatocytic origin of both cell lines and confirmed their purity. Although morphologically similar to normal primary hepatocytes, RNT cells were able to survive and grow in monolayer culture for months and were not tumorigenic in vivo. On the contrary, RH cells displayed tumor-initiating cell markers, formed numerous colonies in soft agar and spheroids when grown in 3D and were highly tumorigenic and metastatic after injection into syngeneic rats and immunocompromised mice. Moreover, RNT gene expression profile was similar to normal liver, while that of RH resembled HCC. In conclusion, the two cell lines here described represent a useful tool to investigate the molecular changes underlying hepatocyte transformation and to experimentally demonstrate their role in HCC development.

KW - CD24

KW - CD90.1

KW - HCC cell lines

KW - Immortalized non-tumorigenic cells

KW - Resistant hepatocyte

UR - http://www.scopus.com/inward/record.url?scp=85014066914&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85014066914&partnerID=8YFLogxK

U2 - 10.18632/oncotarget.14984

DO - 10.18632/oncotarget.14984

M3 - Article

C2 - 28157710

AN - SCOPUS:85014066914

VL - 8

SP - 15716

EP - 15731

JO - Oncotarget

JF - Oncotarget

SN - 1949-2553

IS - 9

ER -