The experimental study of 125I-FIAU uptake in myocardial cells infected with Ad-TIV165

Objectives To study the feasibility of Ad-CMV-HSV1-sr39tk-IRES-VEGF165(Ad-TIV165) as a myocardial reporter gene imaging agent in vitro.

Methods Different infection titer of recombinant adenovirus Ad-TIV165 were used to infect myocardial cell cultured in vitro (Ad-EGFP as control group), and the uptake of myocardial cells to probe 125I-FIAU at different times were measured. The expression of HSV1-TK and VEGF165 were detected using RT-PCR and Western-blot.

Results After 24 hours of transfected Ad-TIV165 (MOI 0 and 50) into myocardial cell, the uptake of 125I-FIAU of myocardial cell increased with the virus titer and also the extension of time, the uptake and infection time and infection titer were positively correlated (R2 = 0.8315); when MOI was from 70 to 100, the uptake and infection time and infection titer were negatively correlated. However, the uptake was extremely low in the control group at any virus titer. When MOI was 0, the reporter gene HSV1-sr39TK and the therapeutic gene VEGF165 couldn’t be detected at mRNA or protein level; When MOI was 50, both HSV1-sr39TK and VEGF165 could be detected in infected myocardial cell at mRNA and protein level.

Conclusions There was obvious uptake of 125I-FIAU after myocardial cell was transfected with Ad-TIV165 and the uptake was relevant to the results of mRNA and Western Blot. This provided a theoretical and experimental basis for further experiments in vivo.

Research Support Supported by 30400176, 30970853, 3083004