| Abstract |
Objective: To establish hepatocellular carcinoma (HCC) cell lines and
provide more pure liver carcinoma cells for HCC studies.
Methods: The carcinoma tissues were obtained from the primary lesion and
metastatic lesion in portal vein of HCC. The carcinoma cells were isolated
by Percoll density gradient centrifuge and cultivated. The cell clones
were then obtained by micro-environmental digestion and cell lines were
established. The growth curves, G band chromosome karyotype, comparative
genomic hybridization (CGH) of the established cells were characterized.
The cells were implanted in nude mice for tumorigenesis.
Results: 20 cases of HCC cells were isolated and cultivated. Two cell
lines (designated as H2M and H4M) were successfully established from 2
cases of emboli of portal vein. The karyotype showed that both cell lines
are a hypertriploid (71~78 chromosomes). A marker chromosome containing 1q
and 6p [t(1;6)] was found in H2M cells and a huge marker chromosome
containing a long homogeneously staining region (hsr) in H4M cells. The
main genetic alterations analyzed by CGH were a high copy number
amplification of 1q, 3q, 5p, 6p, 7q and 8q and loss of 4q, 13q, 16q, 17p
and 19p in H2M cells, whereas, amplification of 6p, 7p, 11p, 11q13 and
loss of 8p, 9, 13q, 16q in H4M cells. Implantation of H2M cells in nude
mice for a month produces typical human hepatocellular carcinoma, but no
tumor for H4M cells.
Conclusions: Percoll density gradient centrifuge and micro-environmental
digestion is an effective method for cloning of primary carcinoma cell and
cell line establishment. Two established HCC cell lines will provide
culture cells for HCC studies. The genetic alterations detected in both
cell lines also provide clues and cell models for further screening of
oncogenes and tumor suppressor genes in HCC.
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