The Study Of Anti-tomor Mechanism Of Seleno-chitosn In Calu-1 Lung Cancer Cell

In this study,human lung tumor cells calu-1 were incubated with different concentrations of Seleno-Chitosn(SC)for 48 h to study the apoptosis mechanism.First,seleno-short-chain-chitosn was found that have the biological activity of promoting the human lung cancer calu-1 cells apoptosis by MTT assay.The results showed that the inhibitory effect of seleno-short-chain-chitosn on the proliferation of calu-1 cells became stronger as the concentration and time of action of seleno-short-chain-chitosn increased,and the inhibitory effect was positively correlated with the action time and concentration.When the concentration was 200 g/mL and co-cultured with calu-1 cells for 48 h,the inhibitory effect on the proliferation of calu-1 cells was significant.Cell viability can be reduced to less than 30%.By scanning electron microscopy,Hoechst 33342/PI et al.found that under the action of seleno-short-chain-chitosn e at different concentrations,calu-1 cells showed typical apoptosis characteristics such as shrinkage,cell membrane rupture and apoptotic vesicles during apoptosis.Annexin v-fitc/PI staining assay further demonstrated that chitosan selenate can induce apoptosis of calu-1 cells,and the degree of apoptosis is related to the action concentration.At the same time,the cycle distribution of calu-1 cells under the action of chitosan selenate at different concentrations was detected by flow cytometry.The results showed that chitosan selenate destroyed the continuity of calu-1 cell cycle and blocked its cell cycle at G2/M and S phases.Second,with concentration of 5 mM n-acetyl-L-cysteine(NAc),as a pretreatment,oxidation inhibitors of cells by flow cytometry detected under the action of different acid concentration of selenium chitosan Calu-1 is the change of intracellular reactive oxygen content with the concentration increase of selenate chitosan and rising,and this change is accompanied by the mitochondrial transmembrane potential of gradually disappear.Therefore,we speculated that chitosan selenate may play a role in inducing apoptosis in the endogenous mitochondrial apoptosis pathway of calu-1.In this study,relative quantitative analysis was performed on the mRNA expression of Bax,bcl-2,caspase-3 and caspase-9,the key genes regulating apoptosis in cells at the transcriptional level,with-actin as the internal reference.It was further verified that chitosan selenate could promote apoptosis of calu-1 cells by regulating the expression of apoptosis-related genes.Finally,at the translation level,the expression differences of related apoptotic proteins were detected by relative quantitative analysis to verify that Seleno-Short-Chain-Chitosncan promote apoptosis of calu-1 cells.Western blotting was used to analyze the expression of apoptosis-related proteins in calu-1 cells.In summary,chitosan selenate has a significant inhibitory effect on the proliferation of calu-1 cells,and its pro-apoptotic effect is carried out through the mitochondrial pathway.Therefore,as an anticancer active substance,Seleno-Short-Chain-Chitosn has great research value and potential.