Dual Aptamer Modified Drug Delivery System Mediated By Recombinant Adenovirus Improves The Therapy Effect In Prostate Cancer

Objective: It was found that aptamer DUP-1(Peptide aptamer)can be targeted to prostate cancer cells with negative prostate-specific membrane antigen(PSMA),and A10-3.2 aptamer(RNA aptamer)can be targeted to PSMA-positive prostate cancer cells.Moreover,tumor-suppressor gene of PTEN(phosphatase and tensin homolog deleted on chromosome ten)and doxorubicin(DOX)can effectively inhibit prostate cancer,and the recombinant adenovirus(Ad5)could mediate high gene transfer efficiency.Based on these discoveries,in this study we managed to design the dual aptamers modified tumor targeting gene and doxorubicin delivery system mediated by recombinant adenovirus(A10-3.2(DOX)/DUP-1-PEG-Ad5,ADDP-Ad5),detecting the cytotoxicity,cellular uptake in vitro,and acute toxicity in vivo of the drug delivery system.Methods: In this delivery system,the dual aptamers of A10-3.2 and DUP-1 were connected with adenovirus by PEG(Polyethylene glycol)using amide reaction principle;tumor suppressor gene of PTEN was integrated into DNA of Ad5 and the chemotherapeutic agent of DOX automatically bound with the double chain RNA of aptamer A10-3.2.Moreover,the cytotoxicity and cellular uptake of drug delivery system were detection by the human normal prostate epithelial cells of WPMY-1,liver cancer cells of HepG2,prostate cancers of LNCaP and PC3 in vitro.Finally,acute toxicity experiment demonstrated the toxicity in vivo.Results: After detecting and calculating we knew the PEG modification rate on capsid protein of adenovirus was 98.7 ± 2.43%,the aptamer DUP-1 and A10-3.2 modified products were in a yield of 80.4±1.36% and 82.2 ± 2.14%,respectively.It also suggested that the best molar ratio of DOX embedded into aptamer A10-3.2 is 1:2.In vitro toxicity experiments,the degree of toxicity effect on cells of ADDP-Ad5 was LNCaP,PC3,HepG2,WPMY-1,sorted by toxicity in descend order,and the cellular uptake of LNCaP was more than PC3's cellular uptake for ADDP-Ad5.The results of acute toxicity test showed that high dose group of ADDP-Ad5 made a slight toxicity on liver and heart in mice.Conclusions: All these results proved the successful preparation of the aimed dual aptamers modified tumor targeting gene and DOX delivery system mediated by recombinant adenovirus.This drug delivery system can significantly inhibited prostate cancer,providing aptamer targeting,and showed a slight toxicity on liver and heart in mice at a certain concentration.