Studies on the Growth and Transfer Behaviour of Hca-F Cell and Immunomodulatory of fucoidan
|School||Dalian Medical University|
|Course||Biochemistry and Molecular Biology|
|Keywords||Fucoidan Shift Immunomodulatory Matrix metalloproteinase|
Objective: Fucoidan, a sulfated polysaccharide extracted from brown seaweed exerts anti-viral, anti-bacterial, anti-tumor effects, anti-oxidant, anti-coagulant and anti-inflammatory. Fucoidan have become the focus of a substantial amount of attention and the hot spot of marine drugs because of its potential biological effect. This experiment will further discuss the effect of fuocidan on the growth and transfer behaviour of hepatocellular carcinoma cell,the immune cells of normal mice and discuss the mechanism of anti-tumor effect.Methods: 1.Acquiring fucoidan sample by precipitating with the different concentrations of ethanol, detecting sugar content, sulfate content and uronic acid content of the sample by sulphoacid-phenol method and gelatin- barium chloride method.2.Inhibition rate of fucoidan on the Hca-F cell line was assessed by MTT assays. we examined the effects of fucoidan on tumor invasion and migration by in vitro adhesion and migration assays.3.The effect of fucoidan on the expression of transfer related proteins were assessed by the gelatin zymography, ELISA and RT-PCR analysis.4.The lymphocyte proliferation was assessed by MTT assays. The effect of fucoidan on phagocytosis of macrophages were studied by neutrol red method. The level of cytokine secretion were detected by ELISA method to react the effect of fucoidan on spleen.Results: 1.Acquired pure fucoidan sample with sugar content rate of 42%, sulfate content rate of 12%.2.Fucoidan can inhibit Hca-F carcinoma growth in vitro but the effect is not apparent. Fucoidan can suppress invasion and migration of carcinoma cells obviously.3.Fucoidan could down-regulate matrix metalloproteinase MMP-2 expression in hepatocellular carcinoma cells at both mRNA and protein levels by Gelatin zymography, Sandwich ELISA and RT-PCR analysis.4.It indicated an increased tendency that the proliferative capacity of lymphocyte, phagocytic activity of macrophages and relative contents of IL-6、IL-10、IL-12、IL-18、TNF-αproduced by fucoidan, in mice spleen.Conclusions: This experimental proof fucoidan immune adjustment and antitumor activity. The mechanism of antitumor may be not only by reinforcing cell mediated, humeral immune responseand accommodating lymphocytesto secrete cytokines but also exert antitumor efficiency through down-regulate matrix metalloproteinase MMP-2 expression and suppress invasion and migration of carcinoma cells.