Characterization of a new rat cell line established from 2' AAF-induced combined hepatocellular cholangiocellular carcinoma

• Published in: In vitro cellular & developmental biology. Animal 2001-01, Vol.37 (1), p.17-25
• Main Authors: Gil-Benso, R, Martinez-Lorente, A, Pellin-Perez, A, Navarro-Fos, S, Gregori-Romero, MA, Carda, C, Callaghan, R, Peydro-Olaya, A, Llombart-Bosch, A
• Format: Article
• Language: English
• Subjects: 2-acetylaminofluorene
• ISSN: 1071-2690; 1543-706X
• DOI: 10.1290/1071-2690(2001)037(0017:COANRC)2.0.CO;2

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.