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.
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1 January 2001
CHARACTERIZATION OF A NEW RAT CELL LINE ESTABLISHED FROM 2′AAF-INDUCED COMBINED HEPATOCELLULAR CHOLANGIOCELLULAR CARCINOMA
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
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In Vitro Cellular & Developmental Biology - Animal
Vol. 37 • No. 1
January 2001
Vol. 37 • No. 1
January 2001
chemical carcinogenesis
c-Met
cytogenetics
HGF
liver tumor
OV-6