The Experimental Study For The Effect Of Spirulina Polysaccharide On Hepatoma Cells

Objective:To investigate the effect of different dosage of polysaccharide from Spirulina platensis(PSP)on the human hepatocarcinoma cell line BEL-7404 cells cultivated in vitro.Explore the possible mechanism of its anticancer effect at molecular level through inhibiting cells proliferation,inducing cells Apoptosis and provide the experimental and theoretical basis for its further development.Methods:Cells cultivated technology in vitro and morphology changed was observant by microscope;the inhibition rate of BEL7404 cells by PSP was detected by flow cytometry;the apoptosis of BEL7404 cells induced by PSP was tested by TUNEL method with flow cytometry.The expression of Bax,Bcl-2,Apaf-1 protein after the human hepatoma cells were treated by PSP was detected by immunocytochemical technology.The activity of Caspase-3 was detected by flow cytometry.Results:①PSP dosage of ranged from 300μg/ml to 1000μg/ml has significant inhibited effect on the proliferation of the BEL7404 cells,dose-dependent manner was found between PSP and cells inhibition.②Inverted phase contrast microscope showed that,opacity in cytoplasm,reduced in volume and weakness in light refractive and cell attachment of cells when the concentration higher than 300μg/ml.The clone growth state was damaged and cells scattered,many cell debris suspension and detachment of cells.③FCM found that,PSP of 300μg/ml and 500μg/ml treated with 24 hours,the cell of apoptotic rate of BEL7404 cell were respectively(20.7±8.1)%and(22.3+9.3)%..④The immunocytochemistry showed that PSP caused the increasing of Bax,Apaf-1 protein of BEL7404,while decreasing of Bcl-2 protein.⑤The result of flow cytometry tested the Caspase-3 cell viability suggested that,the viability of Caspase-3 strengthen with the concentration of PSP,the higher concentration,the greater viability.Conclusions:①PSP has cytotoxicity effect on BEL7404 cells and induce cells apoptosis.②The cells apoptosis mechanism of BEL7404 cells induced by PSP is possible with the pathway of mitochondria or death receptor.By activating Caspase-3,the expression of Bax,Apaf-1 protein increased while the Bcl-2 protein reduced and induce cells apoptosis.③PSP may be the potential drug to treat hepatic carcinoma.