Mitochondrial Pathway Research Of SGC-7901 Induce Apoptosis By Oridonin

Oridonin is the main constituents of rabdosia rubescens,which has good antitumor activity,this paper primarily through experiment to research on inducement of Orido-nin on inhibit prolife-ration and apoptosis in SGC-7901 cell and its mechanicsms.The experiments using the MTT assay Cytotoxicity of Oridonin on SGC-7901 cells.In research we uesd inverted microscope,AO staining SGC-7901 for fluorescence microscopy of cell morphology changes,PI staining flow cytometry detection of apoptosis rate.Fluo-3/AM staining CLSM detection the effect of oridonin on Ca2+ concentration in SGC-7901 cells.JC-1 staining CLSM detection the effect of Oridonin on ??m in SGC-7901 cells.FCM detection the effect of Oridonin on Cytochrome C in SGC-7901 cells.Caspase-3 activity assay kit detection the effect of Oridonin on Caspase-3 activity in SGC-7901 cells.IC50 of oridonin to SGC-7901 cell was 22.74 ?mol/L.After 24h of different doses of oridonin for human gastric cancer cell line SGC-7901,inverted microscope observation of negative control group growth of cell adhesion,cell membrane integrity.Positive control group 18 ? mol/L hydroxy-camptothecin following administration of cell surface wrinkling most cell disruption,cell morphology is not complete.In each group,higher concentration of oridonin made the density of cell growth becomes sparse,wrinkles on the surface.In the high concentration(22 ?mol/L)groups,most of the cell,cells incomplete,fewer adherent cells.AO dye fluorescence microscope observation of SGC-7901 cell morphology,cell negative group round,intact,no broken and wrinkle.In each group,higher concentration of oridonin made the density of cell growth becomes sparse,wrinkles on the surface.In the high concentration(22 ? mol/L)groups,most of the cell,cells incomplete.Live nucleus is yellow-green fluorescence,Ms red fluorescence.Apoptosis of proliferating cell nuclear chromatin is yellow-green thick nuclear membrane inside,visible membrane bubbly fungus.Flow cytometry analysis of SGC-7901 apoptosis rate,extended and elevated doses over time,apoptosis rate gradually increased.Results from the three reach the following conclusion:oridonin is able to induce SGC-7901 apoptosis.Different doses(1.25,2.5,5 mol/L)of oridonin makes Laser Confocal scanning microscope inspection found SGC-7901 elevated intracellular calcium ions,and has a measure of dose-dependent after 24H.Study on SGC-7901 cell mitochondrial membrane potential found that oridonin can reduce cell mitochondrial membrane potential and has a measure of dose-dependent;on SGC-7901 cells in the Cytochrome c of the study found that the oridonin Cytochrome C can increase cell concentrations;Caspase-3 Kit for detecting Caspase-3 enzyme found in the cell,oridonin can significantly increase the activity of SGC-7901 cell Caspase-3 enzymes.In terms of mechanism,oridonin is able to increaseCa2+ concentrations in SGC-7901 cells;Ca2+ concentration changes can trigger mitochondrial permeability pore opening,reduce mitochondrial membrane potential,change mitochondrial permeability,induce mitochondrial release cytochrome C,Caspase-3 and Caspase-9Through experiments,we reach the following conclusion:oridonin have certain anti-tumor activity on SGC-7901 and induce apoptosis in mitochondrial pathway.