Abstract:
Objective To construct the recombinant adenovirus vector containing the human prostate-specific membrane antigen (PSMA) gene; and to check the target gene expression on dendritic cells (DCs).
Methods Primers containing the restrictive endonuclease Sfi I were designed, and a full-length PSMA cDNA was obtained from the pCMV-SPORT6 /PSMA plasmid via polymerase chain reaction (PCR). The PCR product was digested with Sfi I and then inserted directionally into pShuttle-CMV-EGFP. The pShuttle-EGFP-PSMA plasmid was then lined with Sfi I. The fragment containing PSMA was reclaimed and transfected into a pAdxsi vector after digestion with I-CeuI and I-SceI. The recombinant adenovirus plasmid was then transfected into human embryonic kidney 293 (HEK293) cells and packed, and Ad -PSMA-green fluorescence protein (GFP) was infected into the DCs from the peripheral blood of healthy volunteers. The PSMA protein expression in the DCs was confirmed via GFP observation and detected using Western blot analysis.
Results The recombined adenovirus PSMA was successfully constructed at a titer of approximately 2× 1010 pfu/mL. PSMA can be expressed efficiently and correctly in DCs after infection.
Conclusion The DC vaccine was successfully infected using the recombinant adenovirus Ad-PSMA. The current results can serve as a good foundation for further research on prostate cancer therapy.