Study of the expression of hytk gene regulated by Egr-1 promotor in human bladder carcinoma cell / 中华泌尿外科杂志

ABSTRACT

Objective To construct radiation inducible-specific hytk gene therapy vector using radiation inducible gene Egr-1 promoter,and to determine its feasibility in bladder carcinoma gene therapy. Methods A radiation inducible-specific hygromycin phosphotransferase-thymidine kinase fusion gene(hytk)vector was constructed by subcloning hytk gene into pCIneo/Egr-1 vector, which was carrying Egr-1 promoter.Then the vector was transferred into bladder carcinoma cells lines EJ with FuGENE TM 6.Antitumor effects were observed after irradiation with 60 Co-rays as well as gancyclovir(GCV) treatment. Results The new cell line,EJ/Egr-1-hytk,incorporated and expressed the hytk gene,which was proved by RT-PCR and Western blotting.Egr-1 promoter actively drived hytk gene expression in the presence of ?-rays in the EJ cells, while lower hytk expression was detected without ?-rays.Antitumor effects were observed after ?-rays irradiation with 60 Co rays and GCV treatment. Conclusions Hytk-based vector harboring Egr-1 promoter is a novel ideal candidate vector for bladder carcinoma gene therapy.