ROSARIO GIL-BENSO, ANTONIO MARTINEZ-LORENTE, ANTONIO PELLIN-PEREZ, SAMUEL NAVARRO-FOS, M. AURELIA GREGORI-ROMERO, CARMEN CARDA, ROBERT CALLAGHAN, AMANDO PEYDRO-OLAYA, ANTONIO LLOMBART-BOSCH
In Vitro Cellular & Developmental Biology - Animal 37 (1), 17-25, (1 January 2001) https://doi.org/10.1290/1071-2690(2001)037<0017:COANRC>2.0.CO;2
KEYWORDS: liver tumor, cytogenetics, chemical carcinogenesis, OV-6, HGF, c-Met
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.