Establishment of cancer cell lines from rat hepatocholangiocarcinoma and assessment of the role of granulocyte-colony stimulating factor and hepatocyte growth factor in their growth, motility and survival.
OBJECTIVE
Oval cells (OCs), putative hepatic stem cells, may give rise to liver cancers. We developed a carcinogenesis regimen, based upon induction of OC proliferation prior to carcinogen exposure. In our model, rats subjected to 2-acetylaminofluorene/ partial-hepatectomy followed by aflatoxin injection (APA regimen) developed well-differentiated hepatocholangiocarcinomas. The aim of this study was to establish and characterize cancer cell lines from this animal model.


METHODS
Cancer cells were cultured from animals sacrificed eight months after treatment, and single clones were selected. The established cell lines, named LCSCs, were characterized, and their tumorigenicity was assessed in vivo. The roles of granulocyte-colony stimulating factor (G-CSF) and hepatocyte growth factor (HGF) in LCSC growth, survival and motility were also investigated.


RESULTS
From primary tumors, six cell lines were developed. LCSCs shared with the primary tumors the expression of various OC-associated markers, including cMet and G-CSF receptor. In vitro, HGF conferred protection from death by serum withdrawal. Stimulation with G-CSF increased LCSC growth and motility, while the blockage of its receptor inhibited LCSC proliferation and migration.


CONCLUSIONS
Six cancer cell lines were established from our model of hepatocholangiocarcinoma. HGF modulated LCSC resistance to apoptosis, while G-CSF acted on LCSCs as a proliferative and chemotactic agent.
Piscaglia
Anna C
AC
Department of Internal Medicine, GI & Liver Stem Cell Research Group, Catholic University of Rome - Gemelli Hospital, Largo A. Gemelli 8, 00168 Rome, Italy. annachiarapiscaglia@hotmail.com
Shupe
Thomas D
TD
Pani
Giovanbattista
G
Tesori
Valentina
V
Gasbarrini
Antonio
A
Petersen
Bryon E
BE
D015073:2-Acetylaminofluorene / Q000633:toxicity; D016604:Aflatoxin B1 / Q000633:toxicity; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D018281:Cholangiocarcinoma / Q000473:pathology*; D016179:Granulocyte Colony-Stimulating Factor / Q000502:physiology*; D006498:Hepatectomy; D017228:Hepatocyte Growth Factor / Q000502:physiology*; D008113:Liver Neoplasms / Q000473:pathology*; D008297:Male; D009362:Neoplasm Metastasis; D019859:Proto-Oncogene Proteins c-met / Q000502:physiology; D051381:Rats; D011916:Rats, Inbred F344; D016188:Receptors, Granulocyte Colony-Stimulating Factor / Q000502:physiology
D016188:Receptors, Granulocyte Colony-Stimulating Factor; D016179:Granulocyte Colony-Stimulating Factor; D017228:Hepatocyte Growth Factor; D015073:2-Acetylaminofluorene; D016604:Aflatoxin B1; D019859:Proto-Oncogene Proteins c-met