Construction of urothelium-specific recombinant adenovirus and its inhibition in bladder cancer cell.
OBJECTIVE
To construct urothelium-specific recombinant adenovirus and investigate its inhibition in bladder cancer cell.


METHODS
RT-PCR analysis was used to determine expression patterns of hUPII and coxsackie adenovirus receptor on multiple cell lines. Transient transfection and luciferase detecting assay were used to detect tissue specificity of the hUPII promoter. Recombinant adenovirus Ad-UPII-E1A and Ad-UPII-Null were constructed. Restrictive enzyme digestion assay and PCR confirmed the correct construction. The adenovirus E1A protein expressed in BIU-87 was tested by Western blot after cells were infected with recombinant adenovirus. Recombinant adenovirus Ad-UPII-E1A was tested for its inhibition in bladder cancer cell line BIU-87.


RESULTS
HUPII and CAR were expressed and the hUPII promoter is highly active in bladder cancer cell line BIU-87. Using homologous recombination in bacteria technology, the hUPII promoter and E1A gene were inserted into the genome of type 5 recombinant adenovirus. The E1A protein was markedly positive in the samples of BIU-87 cells infected with recombinant adenovirus Ad-UPII-E1A. MTT assay demonstrated recombinant adenovirus Ad-UPII-E1A inhibited bladder cancer cell BIU-87 growth.


CONCLUSIONS
The hUPII promoter shows high tissue specificity. Recombinant adenovirus Ad-UPII-E1A and Ad-UPII-Null were constructed and confirmed. Recombinant adenovirus Ad-UPII-E1A is effective in inhibition in bladder cancer cell line BIU-87.
He
Xiang-Dong
XD
Institute of Urology, Second Hospital of Lanzhou University, Lanzhou, PR China.
Wang
Zhi-Ping
ZP
Wei
Hai-Yan
HY
Zhou
Qin
Q
Wang
De-Gui
DG
Tian
Jun-Qiang
JQ
Fu
Sheng-Jun
SJ
Rodriguez
Ronald
R