Establishment And Characterization Of Human Lymphatic Metastatic Hepatocellular Carcinoma Cell Models And Study On The Mechanism Of Organ-specific Metastasis

Hepatocellular carcinoma ( HCC) is the secondary cancer "killer" in China,which accounted for 18.8% of total deaths with malignant diseases, and 53%of all HCC deaths worldwide. Despite of the progress in surgery of liver cancer,the overall prognosis of HCC is still poor. Only 5% of patients with HCC live out5 years, and 60~70% patients occur metastasis or recurrence within 5 yearsafter curative resection. Metastatic recurrence is still the main obstacle to theimprovement of treatment efficacy. Therefore, studies on metastasis andrecurrence are still important issues in the 21st century. Animal models and cell models are essential for the study of mechanismof HCC metastasis. Many animal models or cell models have been establishedand used, but most of them can not form metastasis. Cell line with definitelymphatic metastatic potential is still not yet reported. Metastasis of cancercells is a series of sequential, non-random and highly selective process, inwhich host microenvironment actively participate in the process. Recently, theresearch on organ-specific metastasis is one of the hotspot of metastasisresearch; however the mechanism is still unclear. It is reported that,chemokines and their receptors play important roles in the invasion, migrationand metastasis of cancer cells. Some chemokines and their receptors werefound to be key molecules in specific cancers. These findings provide newtargets or strategies in prevention and treatment. However, there are no reporton the roles of chemokines and receptors on HCC. In the current study, firstly, we used the method of in vivo consecutiveselection to establish the HCC cell line with high lymphatic metastatic potential,called HCCLYM. HCCLYM cells were subcloned using limited dilution method,and two monoclonal strains with different metastatic potentials to local lymphnodes, HCCLYM-H (100%) and HCCLYM-L (30%), were selected. Detect thebasic biological characteristics and the metastatic potentials of the establishedcell line and monoclones. Secondly, the different gene expression profiling ofHCCLYM-H and HCCLYM-L was assessed by chemokines and receptors gene 6博士论文 英文摘要array, to study the roles of chemokines and their receptors in theorgan-specific lymphatic metastasis of HCC, and find the specific chemokinesor chemokines receptors related with organ-specific lymphatic metastasis ofHCC, then further discuss the mechanism of the effects. Part OneEstablishment and characterization of humanlymphatic metastatic hepatocellular carcinoma cellmodels The objective of this part is to establish a HCC cell line with high lymphaticmetastatic potential, then subclone and select two monoclonal strains with thedifferent metastatic potentials, and to detect the basic biologicalcharacteristics and the metastatic potentials of the established cell line andmonoclones. Use the human HCC cell line HCCLM6, which has high metastaticpotentials to multiple directions, as parental cell line. Inject HCCLM6 cells intothe footpads of nude mice, and proliferate the tumor cells in the metastaticlymph nodes by primary culture, then repeat the in vivo selection for 8 rounds.Proliferate and purify the tumor cells in the parental cell line which havespecific high metastatic potential to lymph nodes stage by stage, finally,establish the HCC cell line HCCLYM, which has high lymphatic metastaticpotential. The lymphatic metastatic rate of HCCLYM cells, when transplanted onliver, was 83%, and the rate of lung metastasis was 40%. The cells showedpolygonal shape and adhesive growth when cultured in vitro. The averagediameter of HCCLYM cells was 38±2μm,doubling time was 31.6 hours, andclone formation rate was 24±4%. The HCCLYM cells were triploid, the numberof chromosomes was 45~68, mainstream...