1. Volume 10, Issue 6, Pages 1051-1058 (December 2004)
Targeting Prostate Cancer with Conditionally Replicative Adenovirus Using PSMA Enhancer 
Sang-Jin Lee, Yanping Zhang, Sang Don Lee, Chaeyong Jung, Xiong Li, Hong-Sup Kim, Kyung-Hee Bae, Meei-Huey Jeng, Chinghai Kao, Thomas Gardner 
Molecular Therapy 
Volume 10, Issue 6, Pages (December 2004)
DOI: /j.ymthe
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

2. FIG. 1
The transcription enhancing activity of PSME. (A) PSME sequence was inserted upstream of TATA in pGL3/TATA. (B) The transcriptional activities of this construct were evaluated in the absence of androgen by transient reporter transfection assay as described under Materials and Methods. All data were normalized by cotransfection of 1 ng of a Renilla luciferase vector, pRL-SV40 (Promega). Results are presented as means ± standard error of three transfections. (C) The luciferase activity of pGL3/PSME/TATA or pGL3/RSV was measured in C4-2 cells in the absence of androgen. Relative luminescence is expressed as the mean ± standard error of the mean of at least three independent experiments.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

3. FIG. 2
The schematic organizations of the recombinant adenoviruses used in this study. PSME controls the expression of E1A, resulting in the selective replication of adenovirus Ad5-PSME-E1a in PSME-positive cells. Ad5-CMV-βgal is a replication-deficient virus due to deletion of the E1 and E3 regions. ITR, inverted terminal repeat.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

4. FIG. 3
E1A protein expression mediated by Ad-PSME-E1a. Androgen-independent prostate cancer cells were harvested 2 days after infection and lysed. Cell lysates were resolved by SDS-PAGE and transferred to PVDF membrane. E1A proteins were detected by Western blotting as described under Materials and Methods.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

5. FIG. 4
Replication of Ad-PSME-E1a virus in several human cancer cells in vitro. Cells (1 × 106) were infected with 5 pfu of wild-type or Ad-PSME-E1a per cell. Viruses recovered from cells and media 2 days later were titered on confluent layers of 293 cells. Results are presented by log10 (titerAd-PSME-E1a/titerwt).
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

6. FIG. 5
Cytotoxicity efficacy of Ad-PSME-E1a on prostate cancer and A549 cells in vitro. Cells (5 × 104) were seeded onto six-well plates and infected with different numbers of pfu per cell as shown. After 5 days, viable cells were counted after staining with trypan blue. Each experiment was carried out in triplicate. The results showed that Ad5-PSME-E1a was cytotoxic to androgen-independent prostate cancer cells (C4-2 and CWR22rv).
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

7. FIG. 6
Treatment of tumor xenografts with recombinant adenoviruses. Tumor xenografts (CWR22rv) were grown sc in athymic nude mice. (A) Tumor size was measured at different time points as indicated after intratumoral injection of Ad5-PSME-E1a (1.7 × 108 pfu) or Ad5-CMV-βgal. Each group included four mice with two tumors per mouse. (B) H&E staining of tumors obtained from mice treated with either Ad5-PSME-E1a or Ad5-CMV-βgal. Original magnification ×20. (C) Immunohistochemical staining with anti-adenovirus antibodies and counterstaining (H&E) were performed. Black arrows indicate stained adenovirus foci. Original magnification ×200.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions