Effect And Mechanism Of Cinnamaldehyde On Esophageal Cancer Cells

Objective:To investigate the effect of cinnamaldehyde on the proliferation,cell cycle and apoptosis of esophageal cancer cells in Eca109 and KYSE30,and to explore its anti-tumor mechanism as a theoretical basis for the development of anti-cancer drug.Methods:1 The proliferation of Eca109 and KYSE30 cells treated with different concentrations of cinnamaldehyde were detected by MTS.2 Flow cytometry detected the different concentrations of cinnamaldehyde inducing the cell cycles and apoptosis rate of esophageal cancer cells Eca109 and KYSE30 cells.3 Effect of different concentrations cinnamaldehyde on the level of the expression of apoptosis-related proteins were detect by western blot for 24 h.4 iTRAQ mass spectrometry detected the differentially expressed proteins of KYSE30 cells between the control group and cinnamaldehyde treatment group,and analyse the differentially expressed proteins in Uniprot database by GO and KEGG.Results:1 MTS assayed showed,the inhibition rate of Eca109 and KYSE30 cells proliferation is on the rise increased after treatment with different concentrations(0,7.5,15,30?g/ml)of cinnamaldehyde for 12,24 and 48 hours,and a dose-dependent relationship can been seen between them(P <0.05).2 The result of flow cytometry analysis indicated that,after treatment with different concentrations of cinnamaldehyde for 24 h,the cell cycles and apoptosis rate of esophageal cancer cells were significantly changed,and the percentage of G0/G1 phase was significantly decreased(P<0.05),G2/M phase cells were significantly increased(P<0.05);with increasing concentration of cinnamaldehyde,the difference was more obvious.3 The result of Western blot indicated that,expression of Bcl2 and Mcl1 protein in Eca109 and KYSE30 cells was significantly lower than that in the control group,the difference was more obvious(P<0.05),but the expression of apoptotic protein Bax was significantly up-regulated(P<0.05);the apoptotic proteases Caspase3 and Caspase9 showed significant cleavage fragments,and the differencere was more obvious(P<0.05).4 ITRAQ mass spectrometry analysis showed that 210 differentially expressed proteins were found in KYSE30 cells compared with control group,and GO analysis and KEGG Pathway analysis were performed on 210 different proteins.In the UNIPROT database,205 proteins were identified and 197 proteins were identified,including 28(13.7%)of the proteins involved in the process of positive and negative regulation of apoptosis,involved in intercellular adhesion proteins;There were 204 proteins(51.2%)involved in the cytoplasmic composition,85(41.5%)of the nuclear components,74(36.1%)of the protein of the cytoplasmic protein,and the like.44(21.5%)of poly-RNA,40(19.5%)of ATP-binding protein and so on.In the KEGG Pathway analysis,it was found that 101 differentially different proteins could be involved in pathways in cancer(12),splice(9),MAPK signal transduction pathway(9),apoptotic pathway(7)and VEGF signal transduction pathway(4).Pathways in cancer involved the most commonly protein,but the apoptotic pathways were the most significant differences in proteins(P <0.001).Conclusion:1 Cinnamaldehyde inhibits the proliferation of KYSE30 and Eca109 cells.2 Cinnamaldehyde can cause G2/M arrest in KYSE30 and Eca109 of esophageal cancer cells,and induce apoptosis.3 Cinnamaldehyde may down-regulate the expression of Bcl2 and Mcl1 protein,up-regulate the proapoptotic protein Bax,and promote the activation of downstream apoptotic proteases Caspase3 and Caspase9,interfere with cell cycle progression and induce apoptosis of esophageal cancer cells.4 Protein mass spectrometry analysised the differential protein of KYSE30 cells in esophageal cancer between the control group and cinnamaldehyde treatment group.The differential protein was analyzed by KEGG,and the differential protein was involved in pathways in cancer,splice,MAPK signal transduction pathway,apoptotic pathway and VEGF And P53 signal transduction pathway.The most common protein species are in pathways in cancer,but the apoptotic pathways are the most significant differences among them.