Establishment Of A New Human Hepatocellular Carcinoma Cell Line And Its Biological Characteristics

Aims: Hepatocellular carcinoma (HCC) is a kind of malignant disease that isdifficult to treat and has a high incidence and a high mortality at present. As theincidence of liver cancer and transfer mechanisms can not be directly on the humanbody, so the establishment of cell lines and animal models of human liver cancer areparticularly important for in vitro or in vivo studies. The established cell lines havesettled a good foundation for study of pathogenesis and treatment of HCC. However,each liver cancer cell has different sources and biological characteristics. Yet thesame cell line could be transformed, so the test results are uncertainty. The data showlimit resource of cell lines can’t satisfy the needs of study. So the purpose of thisproject is to develop a new cell lines derived from primary liver cancer, and can beused to study the common and different points of pathogenesis, progress andmetastasis of liver cancer.Methods: The sample of human HCC was primary digested by collagenase. Thesingle HCC cell was obtained by expanding the cell clones. The cells were passagedwith trypsin. Growth status was observed under light microscope. The survival ratewas measured by counting cells stained with protamine blue. Seeding efficiency andcell growth curve was measured by counting cell number. The expression of livermarkers such as AAT、ALB was detected by PCR and Western-blot. Studies of cellcycle were performed by flow cytometry. Xenograft was performed by inoculatingHLCZ02cells into the flanks of the scid mice and the tumors were observed, thusxenograft rate was counted. Chromosomes was made by traditional method andanalyzed by photographed. Cell contamination was detected by DAPI nuclear staining.The cells were infected with HCVcc and intracellular HCV RNA was detected by PCR.We used anticancer drugs TRAIL and sorafenib to treat the cells, and detectedapoptosis using flow cytometry.Results: A new human hepatoma cell line, named HLCZ02, was successfullyestablished. HLCZ02cells grow rapidly in vitro, with the AAT and ALB proteinexpression. Cell cycle analysis showed that DNA synthesis strong, highly malignantcells hypotetraploid. The cells were inoculated subcutaneously into SCID mice andcaused immunodeficient mice tumorigenicity. HLCZ02cells can be infected by HCVand the cells are sensitive to TRAIL and sorafenib treatment. Conclusion: The established HLCZ02cell line maintains the biologiccharacteristics of hepatocellular carcinoma, and might be used to explore thehepatocarcinogenesis and develop antitumor drugs.