Title: Construction of an adenovirus vector driven by human telomerase reserve transcriptase gene promoter and Its expression in bladder carcimona
Abstract: Objective To construct an EGFP labeled recombinant edenovirus vector driven by human telomerase reserve transcriptase (hTERT) promoter and study its expression in bladder carcinoma cell fine 253J.Methods The shuttle plasmid pDC315-Tp-EGFP which contained the hTERT promoter was generated.By using AdEasy system,the replication-defective adenovirus Ad-hTERT-EGFP was assem- bled.Bladder cancer cell line 253J and human lung fibroblast cell line MRC-5 were transfected by the re- combinant adenovirus of 100 multiplicities of infection (MOI).RT-PCR and fluorescence microscopy were used to detect the mRNA and protein expression,respectively.The recombinant edenovirus Ad-EGFP with mCMV promoter served as positive controls.Results The Ad-hTERT-EGFP was generated successfully and its titer was 3.8×10~(10) pfu/mL.The EGFP protein was highly expressed in (85.2±3.3)% 253J cells which were transfected with Ad-hTERT-EGFP,but not in MRC-5 (P0.01).While transfected with the Ad-EGFP,they expressed the EGFP protein in (87.1±2.2)% 253J and (92.5±3.1)% MRC-5 cells respectively (P0.01).By RT-PCR,EGFP mRNA could be detected in both cell fines transfected by Ad- EGFP and 253J cells transfected by Ad-hTERT-EGFP,but couldnt be detected in MRC-5 cells transfected by Ad-hTERT-EGFP.Conclusion The downstream gene controlled by hTERT promoter in adenovirus system can be expressed selectively in bladder carcinoma cell line 253J,which showed the obvious targe- ting character.
Publication Year: 2007
Publication Date: 2007-01-01
Language: en
Type: article
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