| Glycolysis is closely related to angiogenesis of gastric cancer.PFKFB3 is a pivotal rate-limiting enzyme in the process of glycolysis,which has strong kinase activity.By inhibiting its activity,it can reduce the rate of glycolysis and restrain angiogenesis in gastric carcinoma.Regulating glycolysis key rate-limiting enzyme regulatory factor(PFKFB3)regulating angiogenesis of gastric cancer is a new target for the treatment it.Ceria nanoparticles(CeONPs)is a widely used rare earth oxide material with structural properties such as self-regeneration and biomimetic activity,as well as the unique advantages of nanomaterials.Ceria nanoparticles can affect tumor angiogenesis by regulating metabolism,which is one of the most significance medical applications,but its potential mechanism remains to be further elucidated.The research on the influence of CeONPs on angiogenesis in gastric cancer SGC-7901 microenvironment can explore a new mechanism for ceria nanoparticles to regulate tumor angiogenesis and provide a basis for the development of newtumor treatment strategies.Objective:1.The supernatant of human gastric cancer cell line SGC-7901 was co-cultured with HUVECs to construct tumor conditioned medium(Tumor Conditioned Medium,TCM)to simulate tumor microenvironment.The effects of SGC-7901 microenvironment on the proliferation,migration,invasion and tube formation of HUVECs,the protein expression of VEGF,HIF-1a,PFKFB3,and theproduction oflactic acid and ATPwerestudied.2.HUVECs induced by TCM of gastric cancer cell line SGC-7901 was treated with ceria nanoparticles.The changes of proliferation,migration,invasion,tube formation,protein expression of PFKFB3,VEGF,HIF-1a and the production of lactic acid and ATP were detected.To explore the effect of CeONPs on angiogenesis in gastric cancer SGC-7901 microenvironment by inhibiting PFKFB3 activity.Methods:1.The cultivate supernate of SGC-7901 was collected and co-cultured with HUVECs to establish gastric cancer SGC-7901 angiogenesis model to simulate the angiogenesis of SGC-7901.2.Filtrating thetreatment concentration ofceria nanoparticles:the best CeONPs treatment concentration was selected by MTTexperiment.3.The effects of ceria nanoparticles on the migration,invasion and tube formation of HUVECs in the microenvironment of gastric cancer cell SGC-7901 were observed by cell scratch,cell invasion and angiogensis experiments.4.Using WB assay to examine the changes of protein expression of PFKFB3,VEGF and HIF-1a in HUVECs microenvironment of SGC-7901 cells treated with ceria nanoparticles.5.Lactic acid detection kit and ATP detection kit were used to detect the effects of ceria nanoparticles on lactic acid and ATP production of HUVECs glycolysis products in gastric cancercell SGC-7901 microenvironment.Results:1.The microenvironment of gastric cancer cell line SGC-7901 was constructed.The results of co-culture showed that the proliferation,angiogenesis,migration and invasion of HUVECs induced by SGC-7901 TCM were significantly enhanced the expression of PFKFB3,VEGF and HIF-1a in HUVECs induced by SGC-7901 conditioned medium was significantly increased at protein level.The metabolic level showed that the production of lactic acid and ATPin HUVECs induced by SGC-7901 tumor conditioned medium increased.2.The results of MTT test showed that:(1)CeONPs in the concentration range of(1-10)ug/ml,CeONPs had no effect on the proliferation of HUVECs,and it is no statistically significant of the difference.(2)compared with the control group HUVECs,the proliferation ability of HUVECs induced by gastric cancer cell SGC-7901 tumor conditioned medium was stronger than that of control group HUVECs after induction of gastric cancer cell SGC-7901 by tumor conditioned medium for 48 hours,and it is statistically significant of the difference.(3)CeONPs in the concentration range of(5,7,10)ug/ml could inhibit the proliferation of HUVECs induced by gastric cancer cell SGC-7901 TCM,and it is statistically significant of the difference.3.The ability of migration,tube formation invasion of HUVECs induced by SGC-7901 tumor conditioned medium was significantly higher than that in HUVECs induced by SGC-7901 TCM.The HUVECs induced by gastric cancer cells SGC-7901 TCM in treated with CeONPs decreased the ability ofmigration,invasion and tube formation,and it is statistically significant of thedifference.4.The results of Western blotting showed that the expression of HIF-1a,PFKFB3 and VEGF in HUVECs induced by SGC-7901 TCM was higher than that in HUVECs(P < 0.05).The expression of HIF-1a,VEGF and PFKFB3,in human umbilical vein endothelial cells induced by SGC-7901 tumor conditioned medium was significantly decreased after CeONPs treatment.5.The results of lactic acid and ATP detection showed that HUVECs and HUVECs induced by SGC-7901 tumor conditioned medium increased the production of HUVECs lactic acid and ATP induced by gastric cancer cell SGC-7901 TCM,it is statistically significant of the difference.The production of lactic acid and ATP in HUVECs induced by SGC-7901 TCM was significantly decreased after CeONPs treatment.Conclusion:1.Co-cultureof SGC-7901 supernatant and HUVECs can simulatetumor angiogenesis to someextent.HUVECs induced by SGC-7901 tumor conditioned medium can increase its proliferation,migration,invasion and tube formation,increasethe protein expression of VEGF,HIF-1aang PFKFB3 and the production of lactic acid and ATP.2.CeONPs can reducethe invasion,proliferation,migration,and tubulation of HUVECs induced by SGC-7901 TCM,and reducethe protein expression of VEGF,HIF-1aand PFKFB3 andthe production of lactic acid and ATP.3.CeONPscan affect angiogenesis in SGC-7901 microenvironment by inhibiting PFKFB3 activity. |
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