Objective      To construct conditionally replicating oncolytic adenovirus regulated by prostate specific promotor rPB-DR(6) with an insertion of cytosine deaminase (CD) gene driven by mCMV promoter

to observe its antitumor effect

and to explore a new method for the treatment of androgen-independent prostate cancer.       Methods      Recombinant oncolytic adenovirus Ad5-rPC was constructed and packaged using molecular biotechniques

identified by PCR and sequencing

purified by CSCI gradient centrifugation

and tested for PFU

titer detection. Expression of E1A and CD proteins in the adenovirus-transfected prostate cancer cells was detected by Western blotting. Antitumor effect of the recombinant adenovirus on prostate cancer cells was tested by CCK-8 assay.       Results      By PCR and restriction endonuclease assay

rPB-DR(6) promoter and CD gene driven by mCMV promoter were inserted into recombinant adenovirus successfully. The titer of the purified recombinant adenovirus Ad5-rPC was 1.1×10

11

pfu/mL. Western blot results showed positive expression of E1A and CD proteins in transfected prostate cancer cells. The titer of the progeny viruses produced by prostate cancer cells which were transfected by Ad5-rPC was greater than 3.0×10

6

pfu/mL. CCK-8 assay and cell counting method confirmed that Ad5-rPC had antitumor effects on both androgen-dependent and androgen-independent prostate cancer cells.       Conclusion      Recombinant oncolytic adenovirus regulated by prostate specific promoter has the ability of reproduction in prostate cancer cells uniquely

and can specifically kill prostate cancer cells.