Research On The Human Liver Cancer HepG-2 Cells Apoptosis Induced By Selenium Astragalus-polysaccharide

This paper is a study of Selenium Astragalus polysaccharides for human liver cancer HepG-2 cells effect,to explore the mechanism of its action.Firstly,the experiment through MTT assay measured Selenium Astragalus polysaccharides for of human liver cancer HepG-2 cell growth influence and IC50 values were calculated.The results showed that Selenium Astragalus polysaccharides for HepG-2 cell proliferation inhibition effect and calculate the IC50 value is 18.01μg/ml.The use of an inverted microscope to observe the HepG-2 cell morphology Change of Selenium Astragalus polysaccharides effected after 48h hours.Observations found that the negative control group HepG-2 cells were tightly packed,quantity,morphology full,no damage;The positive control group,HepG-2 cells were very few,cells have been damaged and can’t see the original cell morphology;With the increase of the dose,the administration group HepG-2 cell gap increasing,the number of cells gradually reduce,the cells became round bright,less adherent cells,or even some of the cells damaged.After Hoechst33258 staining,observed under a fluorescence microscope,the nucleus of the negative control group was dyed blue uniform size,tightly packed cells,lot of quantity,no damage,no apoptotic bodies;Positive control group nuclear solidified and deformed,cell gap is large,cell disruption,appeared apoptotic bodies;With the dose increases,the nucleus of the administration group solidification phenomena more obvious,and some even shrunk becomes blue highlights,the cell gap becomes larger,the cells deform more and more obvious.After administration of Selenium Astragalus polysaccharides 48h,after staining with propidium iodide,using flow cytometry detect HepG-2 cells,the results of the apoptotic rates were:Selenium Astragalus polysaccharides 10μg/ml group(14.27± 1.07)%,Selenium Astragalus polysaccharides 20μg/ml group(24.70 ± 0.84)%,Selenium Astragalus polysaccharides 40μg/ml group(5.64±0.79)%.Each group compared with the control group P<0.01,statistically significant,indicating that,with increasing doses,apoptosis peak is increases.The above experiment shows Selenium Astragalus polysaccharides can inhibit of human liver cancer HepG-2 cell growth,its causes associated with apoptosis pathway.Then,using laser scanning confocal detect changes of intracellular calcium fluorescence intensity after 48h Selenium Astragalus polysaccharides action,the result is:Selenium Astragalus polysaccharides 3.75μg/ml group 8.58 ± 0.64,Selenium Astragalus polysaccharides 7.5μg/ml group 9.97± 0.80,Selenium Astragalus polysaccharides 15μg/ml group 15.77±1.31(each group compared with the control group P<0.01,statistically significant).Using laser scanning confocal detect changes of Mitochondrial membrane potential fluorescence intensity after 48h Selenium Astragalus polysaccharides action,the result is:Selenium Astragalus polysaccharides 3.75μg/ml group 31.26±1.54,Selenium Astragalus polysaccharides 7.5μg/ml group 23.62±1.57,Selenium Astragalus polysaccharides 15μg/ml group 9.28±0.64(each group compared with the control group P<0.01,statistically significant).Using laser scanning confocal detect changes of Reactive oxygen species fluorescence intensity after Selenium Astragalus polysaccharides action,the result is:Selenium Astragalus polysaccharides 3.75μg/ml group 3.44±0.23,Selenium Astragalus polysaccharides 7.5μg/ml group 4.82±0.58,Selenium Astragalus polysaccharides 15μg/ml group 8.43±1.17(each group compared with the control group P<0.01,statistically significant).These results suggest that the Selenium Astragalus polysaccharides effect on human liver cancer HepG-2 cells,resulting in HepG-2 intracellular calcium concentration,mitochondrial membrane potential Lower,increased levels of reactive oxygen species,resulting in mitochondrial membrane transition pore opening,protons and water a large number of molecules into the mitochondria,resulting in mitochondrial rupture,resulting in apoptosis.Therefore,this paper through the experiment described that Selenium Astragalus polysaccharides inhibited human liver cancer HepG-2 cells added,the mechanism is through the mitochondrial pathway to caused HepG-2 cells Apoptosis.