C-Jun Mediatedautophagy And Radiosensitivity Of Nasopharyngeal Carcinoma Cell Via PI3K/Akt-mTOR Signaling Pathway

Part ? Expression of c-Jun in nasopharyngeal carcinoma and its relationship with clinical prognosisObjectiveTo investigate whether c-Jun is associated with clinicopathological parameters,the relationship between c-Jun expression and prognosis,and the relationship between PI3K,AKT and mTOR expression in nasopharyngeal carcinoma.Methods1.Collect 69 cases of nasopharyngeal carcinoma and 20 cases of normal nasopharyngeal mucosa in clinical patients.2.Immunohistochemistry was used to detect c-Jun expression in tissues and normal mucosa of patients with nasopharyngeal carcinoma.Results1.The expression of c-Jun in normal nasopharyngeal group was significantly lower than that in nasopharyngeal carcinoma,the difference was statistically significant(p<0.05).2.Combined with clinical case parameter analysis,c-Jun was not associated with age,gender,tumor stage(T),lymph node metastasis(N)and distant metastasis(M)in 69 patients with nasopharyngeal carcinoma.3.Survival analysis showed that patients with nasopharyngeal carcinoma with high expression of c-Jun had less overall survival and fewer patients.ConclusionsThe overall survival time of patients with nasopharyngeal carcinoma with high expression of c-Jun is lower than that of patients with nasopharyngeal carcinoma with low expression of c-Jun,and c-Jun is an independent influencing factor for the prognosis of nasopharyngeal carcinoma.Part ? c-Jun mediates radioresistance of nasopharyngeal carcinoma CNE-2R cells via PI3K/AKT-mTOR pathwayObjectiveTo investigate whether c-Jun regulates the radioresistance of CNE-2R cells through the PI3K/AKT-mTOR pathway.Methods1.Bioinformatics software predicts the relationship between target genes of PI3K,AKT and mTOR and c-Jun;2.The dual luciferase reporter gene system verifies that c-Jun regulates the activity of PI3K and AKT promoters to activate the pathway;3.The cells were divided into 2R group,c-Jun-shRNA group,LY294002 group,RAPA(rapamycin)group,interference pathway node gene,Western blot detection c-Jun,PI3K,p-PI3K,AKT,p-AKT,m-TOR,p-mTOR expression.Results1.Bioinformatics software predicts that c-Jun has complementary binding sites to PI3K and AKT genes,and has no binding site to mTOR.PI3K and AKT are target genes of c-Jun.2.Successfully constructed overexpression vector plasmids G V23 8-mut-PI3 KCG,G V23 8-mut-AKT1 plasmid and empty plasmid GV238-NC,respectively,with C-Jun overexpression vector plasmid GV230-mut-c-Jun,empty plasmid GV230-NC was co-transfected into CNE-2R cells,and the double luciferase reporter system showed that the fluorescence activity of the co-transfected PI3KCG overexpression vector plasmid and GV230-mut-c-Jun group was significantly enhanced compared with the control group(P<0.05).The fluorescence activity of transfected GV238-mut-AKT1 and GV230-mut-c-Jun groups was also significantly enhanced(P<0.05).Compared with 2R group,the expression of c-Jun gene,p-PI3K/PI3K?p-AKT/AKT and p-mTOR/mTOR was up-regulated,the difference was statistically significant(P<0.05).Compared with 2R group,LY294002 group inhibited PI3K/AKT After phosphorylation,the expressions of p-PI3K/PI3K?p-AKT/AKT and p-mTOR/mTOR were down-regulated.Compared with 2R group,the expression of p-mTOR was down-regulated in RAPA group,the difference was statistically significant(P<0.05),There was no significant difference in p-PI3K/PI3K and p-AKT/AKT expression(P>0.05).Conclusionsc-Jun can directly target PI3K,AKT,PI3K and AKT are target genes of C-Jun.c-Jun may regulate autophagy in CNE-2R cells through the PI3K/AKT-mTOR pathway.Part ? Effect of c-Jun on radiosensitivity of nasopharyngeal carcinoma CNE-2R xenografts in nude mice via PI3K/AKT-mTOR pathwayObjectiveTo investigate the effects of interfering C-Jun and PI3K/AKT-mTOR pathways on radiosensitivity of nasopharyngeal carcinoma cell line CNE-2R cells in nude mice xenograftsMethodsNude mice were divided into 2R OGy group,2R+10Gy group,C-Jun-shRNA+10Gy group,rapamycin+l OGy inhibition group,LY294002+10Gy group,IGF-1+10Gy group,and the third group was inoculated with c-Jun silenced CNE-2R cells,the other groups were inoculated with CNE-2R cells,the first group as the control group did not receive irradiation and other treatment,the other groups received 10Gy irradiation dose,rapamycin+10Gy inhibition group,LY294002+1 OGy group,IGF-1+10Gy group were given intraperitoneal injection,IGF-1,LY294002,rapamycin.The volume growth curve of nude mice xenografts was measured,and the expressions of Western blot pathway proteins c-Jun,PI3K,p-PI3K,AKT,p-AKT,m-TOR,p-mTOR,LC3?,and P62 were used.Resuls1.Compared with 2R group,the expression of c-Jun,p-PI3K/PI3K,p-AKT/AKT and p-mTOR/mTOR in 2R+10Gy group was decreased,the difference was statistically significant(P<0.05).2.Compared with 2R+10Gy group,the expression of p-PI3K/PI3K,p-AKT/p-AKT and p-mTOR/mTOR increased in c-Jun-shRNA+lOGy group and IGF+10Gy group,the difference was statistically significant.(P<0.05).The expression of p-PI3K/PI3K p-AKT AKT and p-mTOR/mTOR was decreased in LY294002+10Gy group and RAPA+10Gy group.ConclusionsAfter radiation,c-Jun autophagy was observed in nude mice transplanted with nasopharyngeal carcinoma CNE-2R cells through PI3K/AKT-mTOR pathway.