The Study On The Molecular Mechanism Of Hepatocellular Carcinoma Pathogenesis From Toll Signaling Transduction Pathway In Dendritic Cells

[Objective] Induce DC from PBMC of the healthy men with granulocyte monocyte colony stimulating factor(GM-CSF) and IL-4, LPS induce DC to be more mature. Identify the maturation degree of DC through different approaches. Provide experimental evidence for LPS inducing the maturation of DC. Detect the TLRs mRNA expression in DC by semi-quantitative RT-PCR. Detect the distribution of NF- κB in the nuclear. Study the mechanism of LPS inducing mature DC from gene and molecular level. Induce HCC DC from PBMC of HBV DNA- positive HCC patients, adding LPS to induce it to be mature. Compare the difference of HCC DC and normal DC from the morphology and function. Analyze the level of TLRs mRNA on HCC DC. Put forward the theory of TLRs signaling transduction pathway defection of HCC patients. Offer new explanation for hepatocellular carcinoma pathogenesis.[Methods] Induce DC from PBMC of the healthy men with granulocyte monocyte colony stimulating factor(GM-CSF) and IL-4. Adding LPS on the day 6 of culture, cultivate for another 24h. Harvest the cells to examine. To observe the morphology of DC by inverted phase contrast microscope, electron microscope and laser scanning confocal microscope; Analyze the phenotype of DC with flow cytometry; Investigate the endocytosis ability of DC as a group by Horseradish peroxidase endocytosis assay; Appraise the allogeneic mixed lymphocytes reaction of DC by MTT reduction assay; Analyze the levels of IL-12,TNF-α in the culture medium of DC and IFN-γ in MLR by ELISA; Decide the maturation degree of DC from the...