1. Volume 54, Issue 3, Pages 621-630 (September 2008)
Celecoxib has Potent Antitumour Effects as a Single Agent and in Combination with BCG Immunotherapy in a Model of Urothelial Cell Carcinoma 
Simon J. Dovedi, John A. Kirby, Barry R. Davies, Hing Leung, John D. Kelly 
European Urology 
Volume 54, Issue 3, Pages (September 2008)
DOI: /j.eururo
Copyright © 2008 European Association of Urology Terms and Conditions

2. Fig. 1
Urinary levels of prostaglandin E2 (PGE2) are elevated in patients with high-grade superficial urothelial carcinoma following the initial instillation with bacillus Calmette-Guérin (BCG). A significant increase in urinary PGE2 was detected after initiation of BCG therapy, when compared to pretreatment levels (*p<0.05 Mann-Whitney test). Urinary PGE2 levels were normalised to urinary creatinine.
European Urology  , DOI: ( /j.eururo )
Copyright © 2008 European Association of Urology Terms and Conditions

3. Fig. 2
Bacillus Calmette-Guérin (BCG) causes a dose-dependent increase in urinary prostaglandin E2 (PGE2) levels in C3h/HeSn mice. Mice were intravesically administered either phosphate-buffered saline or 1×106 or 1×107cfu BCG. Urine samples were pooled at 24 and 48h after treatment with BCG. Treatment with 1×106cfu BCG caused a nonsignificant increase in urinary PGE2 levels; this dropped significantly by 48h (p<0.05). Following intravesical administration of 1×107cfu BCG, significant increases in urinary PGE2 levels were observed at 24h (p<0.001), which by 48h had fallen to levels observed in the untreated controls. Data are expressed as mean±SEM and are representative of groups of five mice.
European Urology  , DOI: ( /j.eururo )
Copyright © 2008 European Association of Urology Terms and Conditions

4. Fig. 3
Bacillus Calmette-Guérin (BCG) dose dependently stimulates dendritic cell (DC) expression of cyclooxygenase 2 (COX-2), prostaglandin E2 (PGE2), and interleukin 10 (IL-10). CD1-derived bone marrow DCs were cultured for 7 d in the presence of granulocyte-macrophage colony-stimulating factor and IL-4. DCs were then stimulated for 72h with either 100ng/ml lipopolysaccharide (LPS) or 5×104 to 4×105cfu BCG/ml. Cells were then lysed in lysis buffer and the expression of COX-2 assessed by Western blotting. COX-2 expression was up-regulated 4.75-fold following coculture with BCG at a dose of 4×105cfu BCG/ml when comparing non-treatment and LPS controls (A). BCG dose dependently stimulates DC production of PGE2 and IL-10 (ANOVA; p<0.001; B and C). Data are expressed as mean±SEM and are representative of at least three experiments.
European Urology  , DOI: ( /j.eururo )
Copyright © 2008 European Association of Urology Terms and Conditions

5. Fig. 4
Intravesical bacillus Calmette-Guérin (BCG) therapy in combination with celecoxib is more effective than BCG alone in an MBT-2 murine model. C3h/HeSn mice were instilled with 5×106 MBT-2 cells for 1h following urothelial disruption with 50μl 0.1N HCl/KOH. Intravesical treatment with 1×107cfu BCG commenced on day 1 (24h after tumour implantation) with a subsequent treatment on day 4. BCG treatment was repeated three times a week for a further 2 wk. Mice receiving celecoxib in combination with BCG were treated with 10mg/kg or 100mg/kg celecoxib twice daily (bid) administered subcutaneously in a volume of 100μl. Mice were sacrificed at day 21. Bladders were weighed and examined to determine tumour burden. Intravesical BCG reduced tumour burden but BCG therapy in combination with 10mg/kg twice a day and 100mg/kg celecoxib twice a day resulted in a significant (p<0.05 and p<0.01, respectively) and dose-dependent reduction in MBT-2 tumour burden (ANOVA p<0.01). Data are expressed as mean±SEM and are representative of groups of 10 mice. Solid line illustrates the mass of a non–tumour-bearing bladder.
European Urology  , DOI: ( /j.eururo )
Copyright © 2008 European Association of Urology Terms and Conditions

6. Fig. 5
Celecoxib enhances tumour infiltration of CD3+ and CD4+ cells in the bladders of MBT-2 tumour-bearing mice when assessed after 21 d. C3h/HeSn mice were instilled with 5×106 MBT-2 cells for 1 h following urothelial disruption with 50μl 0.1N HCl/KOH. Mice were treated with 10mg/kg and 100mg/kg celecoxib twice a day (bid) administered subcutaneously in a volume of 100μl. Mice were sacrificed at day 21 and bladders were frozen for analysis by immunohistochemistry. Three representative bladders were assessed each at three levels. Data expressed as mean±SEM (HPF=high-power field). In all mice CD3+, CD4+, and CD8+ cells were evident in the submucosa. Notably, numbers of CD3+ and CD4+ cells were significantly higher in BCG-treated mice with contrastingly fewer CD8+ cells detected.
European Urology  , DOI: ( /j.eururo )
Copyright © 2008 European Association of Urology Terms and Conditions

7. Fig. 6
Dose-dependent reductions in MBT-2 tumour burden are observed following treatment with twice daily 10 and 100mg/kg celecoxib. C3h/HeSn mice were instilled with 5×106 MBT-2 cells for 1h following urothelial disruption with 50μl 0.1N HCl/KOH. Mice were treated with 10mg/kg and 100mg/kg celecoxib twice a day (bid) administered subcutaneously in a volume of 100μl. Mice were sacrificed at day 21 and bladders were weighed and examined to determine tumour burden. Treatment of mice with twice daily 100 but not 10mg/kg celecoxib significantly reduced MBT-2 tumour burden at 21 d (p<0.01) (ANOVA p<0.05). Data are expressed as mean±SEM and are representative of groups of 10 mice.
European Urology  , DOI: ( /j.eururo )
Copyright © 2008 European Association of Urology Terms and Conditions

8. Fig. 7
No increase in T-cell infiltration in the bladders of MBT-2 tumour bearing mice following treatment with 20 or 200mg/kg celecoxib twice daily was observed at 21 d. C3h/HeSn mice were instilled with 5×106 MBT-2 cells for 1h following urothelial disruption with 50μl 0.1 N HCl/KOH. Mice were treated with 10mg/kg and 100mg/kg celecoxib twice daily (bid) administered subcutaneously in a volume of 100μl. Mice were sacrificed at day 21 and bladders were frozen for analysis by immunohistochemistry. Three representative bladders were assessed each at three levels. Data are expressed as mean±SEM.
European Urology  , DOI: ( /j.eururo )
Copyright © 2008 European Association of Urology Terms and Conditions