The Therapy Of Experimental Glioma With P53 Introduced By An Oncolytic Adenovirus Regulated By Dual Specific Promotors

Back ground:The oncolytic adenovirus therapy is prospective in recent years because it enjoyings all the advantages of immunotherapy,biotherapy and gene therapy.Some researchs showed that oncolytic adenovirus,especial the self-replicated ones,are effective to treat tumors.The anti-tumor gene could be expressed and the functional proteins will the cells.Tumor cells could be restrained or even killed by the protein expressed by the anti-tumor gene.But when the function of oncolytic virus has not specific to tumor cells,it could cause the reverse responses in the body.So the specifility is crucial for the success of oncolytic virus if it want to become a feasible and effective measure for tumor therapy.To establish the oncolytic adenovirus for glioma,we designed the 2 specific promoters to regulated 2 target genes in a novel oncolytic adenovirus Ad-Tp-E1A-EnGp-P53.The 2 genes,E1A and P53,entitled the oncolytic adenovirus the power of self replicating and anti-tumor.After that,the specific and effectivity of the onconlytic adenovirous Ad-Tp-E1A-EnGp-P53 was verified and tested in vivo and in vitro.Objective:To assemble the oncolytic adenovirus Ad-Tp-E1A-EnGp-P53 and to verify and test the effectivity and speciality of the oncolytic adenovirus for the treatment of glioma.Methods:Several plasmieds were constructed by means of genetic recombination technology.The conditional replicative recombinant adenovirus Ad-Tp-E1A-EnGp-p53 was assembled successful.The role of anti-tumor of the oncolytic adenovirus was evaluated by MTT and TCID50,with the control virous of Ad-CMV-EGFP.The titer and the growth inhibiting of Ad-Tp-E1A-EnGp-p53 for U251 cells also detected.The expression of P53 and Caspase 3?Cleveaed–Caspase3?in U251 cell were evaluated after the therapy of Ad-Tp-E1A-EnGp-p53.Ad-Tp-E1A-EnGp-p53 also used to treat the nude mice model with glioma.The PET-CT scan was performed for the gliomas in nude mices every 4 days after treat with Ad-Tp-E1A-EnGp-p53.The SUV volume in tumor area was measured.After the nude mice were executed,ERCB expression in tumors of nude mice were evaluated.Results:?1?Theconditionalself-replicatedoncolyticadenivirus Ad-Tp-E1A-EnGp-p53 is assembled successful.The adenovirous can restain the growth of or even kill glioma cells,meanwhile,is irrelevant to normal cells.?2?Western blot showed P53 protein level increased after the treatment of U251 with Ad-Tp-E1A-EnGp-p53?p<0.05?,and there may be the dose-effect relationship.?3?The high expression of Caspase-3 and Cleaved-Caspase-3 can be induced by the Ad-Tp-E1A-EnGp-p53 in U251 cells?p<0.05?.?4?The glioma growth in nude mice can be restrained by Ad-Tp-E1A-EnGp-p53 and the CREB expression in the tumor decreased at the same time?p<0.05?.PET-CT scan showed the decrease of ¹⁸F-FDG intake rate after the treatment of Ad-Tp-E1A-EnGp-p53 in nude mice with glioma.Tumor volume and SUV value decreased after Ad-Tp-E1A-EnGp-p53 therapy?p<0.05?.Conclusion:It is the first time to successfully establish the novel oncolytic adenovirus Ad-Tp-E1A-EnGp-p53 regulated by hTERTp and EnGFAP promoters.Ad-Tp-E1A-EnGp-p53 is effective and specific in vivo and in vitro for glioma therapy.The oncolytic adenovirus could be a new option in comprehensive therapys for glioma.