Characterization of a New Rat Cell Line Established From 2′aaf-Induced Combined Hepatocellular Cholangiocellular Carcinoma

Abstract

A rat cell line-nominated CC-62 derived from a combined hepatocellular and cholangiocellular carcinoma obtained by administration of 2-acetylaminofluorene to male Wistar rats, has been established. Using light and electron microscopy it was determined that morphologically the tumor consisted of a mixed population of hepatocytes and cholangiolar neoplastic cells, intermingled with small, undifferentiated oval-like cells. The CC-62 line has been maintained through 90 passages in culture adopting a paving stone arrangement. Doubling time at the 12th passage was 23 h. Immunostaining with a panel of antisera was performed to identify the cytological profiles of the cell line. There was no k-ras or p53 expression by immunohistochemistry, and molecular biology failed to detect mutations. Molecular analysis by reverse transcriptase-polymerase chain reaction revealed transcripts for c-met but no expression of HGF messenger ribonucleic acid. Three cell lines cloned from CC-62 showed the same immunohistochemical and molecular pattern as the parental line. Cytogenetic analysis revealed a chromosome number ranging from 74 to 82 with a modal number of 79 but no clonal structural abnormalities were found. Deoxyribonucleic acid ploidy analysis showed an aneuploid peak. CC-62 caused tumors 1 mo after subcutaneous transplantation into nude mice, with morphological patterns of mucosecretory solid and spindle-shaped carcinoma. This cell line is the first established from a primary rat combined hepatocellular and cholangiocellular neoplasm. The resulting cells expressed biological and morphological markers of hepatocytes and cholangiolar cells. Therefore this cell line may contribute to a better understanding of the histogenesis of liver cancer.

Cholangiocarcinoma: Molecular targeting strategies for chemoprevention and therapy

“…2,7 A hepatic stem cell origin of intrahepatic cholangiocarcinoma also has been proposed, 12 but not as yet proven. However, the histogenesis of intestinal-type cholangiocarcinomas and of combined hepatocellular and cholangiocarcinoma observed in experimental rodent models of liver carcinogenesis 13,14 and in humans 15,16 is consistent with the concept that at least some subtypes of cholangiocarcinoma may be derived from a pluripotent liver stem cell.…”

Prospero-related homeobox 1 (Prox1) is a stable hepatocyte marker during liver development, injury and regeneration, and is absent from “oval cells”

“…9, 10, Table 2) might also represent cells for carcinogenic predisposition, which was found by Gil-Benso et al (2001), who established an intermediate hepatocellular-/cholangiocellular carcinoma cell line from the AAF/PH rat liver regeneration model. The cell line showed markers shared with hepatocytes and biliary epithelial cells.…”

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

“…Typically, AFB 1 administration in rats results in HCCs arising from hepatocytes, although it might also give rise to CCC [19]. Cyclic feeding with high doses of 2AAF has been shown to induce neoplastic nodules in rats [20]. However, in our model, control rats did not develop liver cancers.…”