Characterization Of Newly Established Human Hepatoma Cell Lines And Experimental Therapy Of TRAIL On Animals Bearing Hepatoma

Primary liver cancer is one of the most common cancers in China, which is an aggressive malignant disease with poor prognosis. It is important to conduct extensive research on its carcinogenesis,biology,prevention and treatment, including the establishment of human hepatoma cell lines and animal models. Recently, six hepatoma cell lines have been established in our institute,and their characters is now carried out for further relative research which constitutes a part of this thesis. In recent years, TRAIL, tumor necrosis factor (TNF)-related apoptosis inducing ligand,a new member of TNF superfamily, has raised great interests among oncologists as it can selectively kill tumor cells while spare non-maligant cells. In our work it was examined whether TRAIL can also induce the apoptosis of the six newly established human heptoma cell lines in vitro and in vivo.Part â… : Characterization of six newly established human hepatoma cell lines. These cell lines have been cultured over 100 passages and have a doubling time between 20 and 60 hours; Microscopic examination showed Morphological features were in consistence with their neoplastic hepatocellular origins. ELISA failed to detected the hepatoma-related proteins in the cell culture supernatants including AFP, albumin, HBsAg, preS. Chromosome analysis showed that: the average numbers of chromosomes of five cell lines were: 51(GBY),56 (KYC),105(LYX),53 (QYC),50( XSM), respectively.Their LDH isozyme spectrums were the same as their clinical counterparts; Cell cycle analysis showed that the ratio of cells in S-phase increased for all the six lines compared with healthy human PBL. Part â…¡: Tumorigenity of the six cell lines. Only subcutaneous injection of KYC and LYX into nude mice led to tumor formation while other cell lines failed to develop tumor. The recultured cell lines from the tumor were found to have properties identical to their original KYC and LYX cell lines.Part â…¢: Cytotoxicity of TRAIL on the six cell lines in vitro and in vivo. In vitro study showed that TRAIL was very effective in inducing the apoptosis of the six cell lines and XSM was the most sensitive one. The sensitivity of the other five cell lines was as the following in a descending sequence: QYC,GBY,KYC,XJC, LYX. Therapeutic effect of TRAIL was examined on animal model, bearing KYC tumor , and it was found that TRAIL treatment inhibited tumor growth. Livers of mice from treatment group were found black and scattered with granules, while no abnormal changes were found in livers from the control mice. Further toxicity experiment of TRAIL was performed with normal Kuming mice and nude mice. The result showed that TRAIL had a significant liver toxicity in Balb/C nude mice. The results suggests that TRAIL may be a useful anti-hepotoma agent but, with significant toxic effect on liver. So there should be further study before it can be useful for clinic.