Characterization of a new rat cell line established from 2'AAF-induced combined hepatocellular cholangiocellular carcinoma.
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.
Department of Pathology, Medical School, University of Valencia, Spain.
R
R
Gil-Benso
A
A
Martinez-Lorente
A
A
Pellin-Perez
S
S
Navarro-Fos
M A
MA
Gregori-Romero
C
C
Carda
R
R
Callaghan
A
A
Peydro-Olaya
A
A
Llombart-Bosch
D015073:2-Acetylaminofluorene; D000782:Aneuploidy; D000818:Animals; D001650:Bile Duct Neoplasms / Q000737:chemistry / Q000473:pathology* / Q000648:ultrastructure; D001653:Bile Ducts, Intrahepatic; D006528:Carcinoma, Hepatocellular / Q000139:chemically induced / Q000473:pathology* / Q000648:ultrastructure; D018281:Cholangiocarcinoma / Q000139:chemically induced / Q000473:pathology* / Q000648:ultrastructure; D004273:DNA, Neoplasm / Q000032:analysis; D011905:Genes, ras; D017228:Hepatocyte Growth Factor / Q000378:metabolism; D007150:Immunohistochemistry; D007621:Karyotyping; D008113:Liver Neoplasms / Q000139:chemically induced / Q000473:pathology* / Q000648:ultrastructure; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008854:Microscopy, Electron; D019859:Proto-Oncogene Proteins c-met / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured / Q000166:cytology* / Q000648:ultrastructure; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
D004273:DNA, Neoplasm; D016159:Tumor Suppressor Protein p53; D017228:Hepatocyte Growth Factor; D015073:2-Acetylaminofluorene; D019859:Proto-Oncogene Proteins c-met