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Updated: Jun 24, 2026
Expression of Transgenes in Native Bladder Urothelium Using Adenovirus-Mediated Transduction
Published on: October 6, 2022
Construction of urothelium-specific recombinant adenovirus and its inhibition in bladder cancer cell
Xiang-Dong He1, Zhi-Ping Wang, Hai-Yan Wei
1Institute of Urology, Second Hospital of Lanzhou University, Lanzhou, PR China.
Aim:
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.
Conclusion:
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.
Insights
Researchers constructed a urothelium-specific recombinant adenovirus (Ad-UPII-E1A) that effectively inhibits bladder cancer cell growth. This targeted approach shows promise for bladder cancer therapy.
Area of Science:
- Oncolytic virology
- Molecular oncology
- Gene therapy
Background:
- Bladder cancer remains a significant health concern, necessitating novel therapeutic strategies.
- Targeted gene delivery systems are crucial for enhancing therapeutic efficacy and minimizing off-target effects in cancer treatment.
Purpose of the Study:
- To construct a urothelium-specific recombinant adenovirus.
- To investigate the inhibitory effect of the recombinant adenovirus on bladder cancer cells.
Main Methods:
- Expression analysis of hUPII and coxsackie adenovirus receptor (CAR) using RT-PCR.
- Tissue specificity of the hUPII promoter confirmed via transient transfection and luciferase assay.
- Construction and confirmation of recombinant adenoviruses (Ad-UPII-E1A and Ad-UPII-Null) using bacterial homologous recombination, restriction enzyme digestion, and PCR.
- Western blot analysis for E1A protein expression in infected BIU-87 cells.
- MTT assay to evaluate the inhibitory effect of Ad-UPII-E1A on bladder cancer cell growth.
Main Results:
- The hUPII promoter demonstrated high activity and specificity in the bladder cancer cell line BIU-87.
- Successful construction of Ad-UPII-E1A and Ad-UPII-Null confirmed by molecular assays.
- Significant inhibition of bladder cancer cell BIU-87 growth was observed following infection with Ad-UPII-E1A.
Conclusions:
- The hUPII promoter exhibits high tissue specificity, making it suitable for targeted gene delivery.
- The constructed recombinant adenovirus Ad-UPII-E1A effectively inhibits bladder cancer cell proliferation.
- Ad-UPII-E1A represents a promising candidate for targeted bladder cancer therapy.
