The Study On Recombinant Adenovirus Mediated E2F-shRNA In Reducing The Growth Of Retinoblastoma

Purpose: To investigate the efficacy of Ad-E2F-siRNA in the process of reducing the growth of Retinoblastoma.Method: The certain amount of Retinoblastoma which has been marked with Green Fluorescent Protein (GFP) cultured in the plate were infected by Ad-E2F-siRNA and Ad-E2F-X respectively. The above infected RB cells will be collected after 48h, then counting the number of cells and diluting them in the concentration of 1*10~5/ml. Injecting the amount of 2*10~5RB in both eyes , the RB tumor cell injected on the left side was infected with Ad-E2F-siRNA as treatment group, simultaneously the right side is infected with Ad-E2F-X as the control group, all of which will be investigated and taken the consecutive and dynamic photos of the condition of RB tumor's growth by the special Fluorescent Microscope within two weeks; At the moment the retinoblastoma tumor cells covers barely all the eyeball, all of the experimental animals, the control group and treatment group will be killed for the following series of experiments, which will be constructive for judging the efficacy of Ad-E2F-siRNA in reducing the growth of Retinoblastoma in terms of molecular and pathological perspective.Results: The results for Realtime RT-PCR showed that we have got the sensitive sequence to the expression of hE2F1, and the amount of tumor cells in the S period of cell cycle was decreased after 48 h infections by hE2F1 siRNA. There is an apparent difference between control group and treatment group investigated by the pathological reactions, in which the area of tumor cells compared to the normal tissues in the control group was larger than the treatment group, the similar result could be captured by the Florescent Scope.Conclusions: The efficacy of Adenovirus-E2F-siRNA is helpful in the process of limiting the growth of retinoblastoma, which can be considered as a new tool for gene therapy.