Prostate stem cell antigen enhancer and uroplakin II promoter based bladder cancer targeted tissue-specific vector

Wang, Degui; Wang, Zhiping; Tian, Junqiang; He, Xiangdong; Chowdhury, Wasim H.; Zhang, Xiangbo; Li, Shigang; Rodriguez, Ronald

doi:10.1016/j.urolonc.2008.02.002

« Previous Next »Urologic Oncology: Seminars and Original Investigations
Volume 28, Issue 2 , Pages 164-169, March 2010

Prostate stem cell antigen enhancer and uroplakin II promoter based bladder cancer targeted tissue-specific vector☆

Degui Wang, Ph.D.
- Institute of Urology, the Second Hospital of Lanzhou University, Lanzhou, China
- School of Basic Medical Sciences, Lanzhou University, Lanzhou, China
Zhiping Wang, Ph.D.
- Institute of Urology, the Second Hospital of Lanzhou University, Lanzhou, China
- Corresponding author. Tel.: +86-931-8942821; fax: +86-931-8942821
Junqiang Tian, Ph.D.
- Institute of Urology, the Second Hospital of Lanzhou University, Lanzhou, China
Xiangdong He, Ph.D.
- Institute of Urology, the Second Hospital of Lanzhou University, Lanzhou, China
Wasim H. Chowdhury, M.S.
- James Buchanan Brady Urology Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA
Xiangbo Zhang, Ph.D.
- Institute of Urology, the Second Hospital of Lanzhou University, Lanzhou, China
Shigang Li, Ph.D.
- Institute of Urology, the Second Hospital of Lanzhou University, Lanzhou, China
Ronald Rodriguez, M.D., Ph.D.
- James Buchanan Brady Urology Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA

Received 4 November 2007; received in revised form 4 February 2008; accepted 5 February 2008. published online 28 April 2008.

Abstract

Purpose

To construct a dual specific vector which contains prostate stem cell antigen enhancer (PSCAE) and uroplakin II (UPII) promoter targeted bladder cancer.

Methods

UPII promoter and PSCAE were amplified by polymerase chain reaction (PCR). Luciferase gene (LUC) was obtained from plasmid pBK-CMV-LUC. PSCAE, UPII promoter and LUC were inserted into shuttle plasmid to create Rp-UPII-LUC and Rp-PSCAE-UPII-LUC. Rp-UPII-LUC and Rp-PSCAE-UPII-LUC were cotransfected with pCMV-β-gal into various cell lines at the presence or absence of androgen receptor agonist R1881 and androgen receptor antagonist flutamide. Luminescence was detected with luciferase assay kit and counted on liquid scintillation counter.

Results

Bladder cancer cells showed higher LUC activity than non-bladder cancer cells after transfected with plasmids Rp-UPII-LUC and Rp-PSCAE-UPII-LUC. PSCAE could improve the LUC activity in both AR positive and AR negative bladder cancer cells but not in non-bladder cancer cells and normal human urothelial (NHU) cells. R1881 could increase the LUC activity in AR positive bladder cancer cells but not in AR negative bladder cancer cells and non-bladder cancer cells. Flutamide could not inactivate PSCAE in bladder cancer cells.

Conclusions

PSCAE can improve target gene expression in bladder cancer cells but not in non-bladder cancer cells and NHU cells. PSCAE maintains a certain level of androgen independent activity in bladder cancer cells. PSCAE is active in both AR positive and AR negative bladder cancer cells. The results suggest that combination of PSCAE with UPII promoter is feasible in constructing bladder cancer-specific vectors.

Keywords: Prostate stem cell antigen enhancer, Uroplakin II promoter, Gene therapy, Bladder cancer