Effect of mitomycin C on concentrations of vascular endothelial growth factor and its receptors in bladder cancer cells and in bladders of rats intravesically instilled with mitomycin C

Avanti Verma, Justin Degrado, Adam B. Hittelman, Marcia A. Wheeler, Hristos Kaimakliotis, Robert M. Weiss

Research output: Contribution to journalArticle

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Abstract

OBJECTIVES • To examine, using in vitro and in vivo models, the largely unexamined effect of mitomycin C (MMC), an effective intravesical treatment for superficial bladder cancer and carcinoma in situ, on expression of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR-2), which mediates many of the angiogenic properties of VEGF. To measure, as a positive control, concentrations of the inhibitor of apoptosis, survivin, as an assessment of MMC effectiveness. To measure MMC-induced changes in proliferation in the presence and absence of VEGF-A small interfering RNA (siRNA). MATERIALS AND METHODS After treatment with increasing MMC concentrations (5-200 Âμg/mL), we measured proliferation, as well as VEGF, survivin, VEGF receptor-1 (VEGFR-1) and VEGFR-2 concentrations in RT-4 and T-24 bladder cancer cells. The effect of pre-treatment of VEGF siRNA and survivin siRNA on MMC-induced decreases in proliferation was measured. Urinary VEGF concentrations and bladder and kidney concentrations of VEGF-A, VEGFR-1, VEGFR-2 and interleukin-6 (IL-6) mRNA were measured in rats intravesically instilled with saline or MMC (200 Âμg/mL). RESULTS Although MMC treatment inhibited cell proliferation and decreased survivin mRNA expression in T-24 and RT4 cells, MMC (12-50 Âμg/mL) increased VEGF-A mRNA and VEGFR-2 mRNA and protein expression. Pre-treatment with VEGF-A siRNA or survivin siRNA before MMC treatment reduced proliferation more than MMC alone. MMC-induced reductions in proliferation were reduced additively by pre-treatment with survivin siRNA, but were potentiated by pre-treatment with VEGF-A siRNA. VEGFR-2 mRNA and protein concentrations and urinary VEGF concentrations were increased in bladders of rats instilled with MMC. CONCLUSIONS Intravesically instilled MMC increases urinary VEGF and bladder VEGFR-2 protein and mRNA in rats. MMC increases VEGF mRNA and VEGFR-2 protein and mRNA concentrations in bladder cancer cells. Therefore, we speculate that MMC could increase the angiogenic potential of both cancer and normal cells. In cancer cells this effect is largest at lower MMC concentrations. Combining MMC with agents that reduce EGF concentrations could be of value in treatment of transitional cell carcinoma of the bladder (TCC).

Original languageEnglish (US)
Pages (from-to)1154-1161
Number of pages8
JournalBJU International
Volume107
Issue number7
DOIs
StatePublished - Apr 2011
Externally publishedYes

Fingerprint

Vascular Endothelial Growth Factor Receptor
Mitomycin
Urinary Bladder Neoplasms
Urinary Bladder
Vascular Endothelial Growth Factor A
Vascular Endothelial Growth Factor Receptor-2
Small Interfering RNA
Messenger RNA
Proteins
Vascular Endothelial Growth Factor Receptor-1
Transitional Cell Carcinoma
Carcinoma in Situ

Keywords

  • bladder cancer
  • intravesical instillation
  • mitomycin C vascular endothelial growth factor
  • vascular endothelial growth factor 2

ASJC Scopus subject areas

  • Urology

Cite this

Effect of mitomycin C on concentrations of vascular endothelial growth factor and its receptors in bladder cancer cells and in bladders of rats intravesically instilled with mitomycin C. / Verma, Avanti; Degrado, Justin; Hittelman, Adam B.; Wheeler, Marcia A.; Kaimakliotis, Hristos; Weiss, Robert M.

In: BJU International, Vol. 107, No. 7, 04.2011, p. 1154-1161.

Research output: Contribution to journalArticle

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abstract = "OBJECTIVES • To examine, using in vitro and in vivo models, the largely unexamined effect of mitomycin C (MMC), an effective intravesical treatment for superficial bladder cancer and carcinoma in situ, on expression of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR-2), which mediates many of the angiogenic properties of VEGF. To measure, as a positive control, concentrations of the inhibitor of apoptosis, survivin, as an assessment of MMC effectiveness. To measure MMC-induced changes in proliferation in the presence and absence of VEGF-A small interfering RNA (siRNA). MATERIALS AND METHODS After treatment with increasing MMC concentrations (5-200 {\^A}μg/mL), we measured proliferation, as well as VEGF, survivin, VEGF receptor-1 (VEGFR-1) and VEGFR-2 concentrations in RT-4 and T-24 bladder cancer cells. The effect of pre-treatment of VEGF siRNA and survivin siRNA on MMC-induced decreases in proliferation was measured. Urinary VEGF concentrations and bladder and kidney concentrations of VEGF-A, VEGFR-1, VEGFR-2 and interleukin-6 (IL-6) mRNA were measured in rats intravesically instilled with saline or MMC (200 {\^A}μg/mL). RESULTS Although MMC treatment inhibited cell proliferation and decreased survivin mRNA expression in T-24 and RT4 cells, MMC (12-50 {\^A}μg/mL) increased VEGF-A mRNA and VEGFR-2 mRNA and protein expression. Pre-treatment with VEGF-A siRNA or survivin siRNA before MMC treatment reduced proliferation more than MMC alone. MMC-induced reductions in proliferation were reduced additively by pre-treatment with survivin siRNA, but were potentiated by pre-treatment with VEGF-A siRNA. VEGFR-2 mRNA and protein concentrations and urinary VEGF concentrations were increased in bladders of rats instilled with MMC. CONCLUSIONS Intravesically instilled MMC increases urinary VEGF and bladder VEGFR-2 protein and mRNA in rats. MMC increases VEGF mRNA and VEGFR-2 protein and mRNA concentrations in bladder cancer cells. Therefore, we speculate that MMC could increase the angiogenic potential of both cancer and normal cells. In cancer cells this effect is largest at lower MMC concentrations. Combining MMC with agents that reduce EGF concentrations could be of value in treatment of transitional cell carcinoma of the bladder (TCC).",
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T1 - Effect of mitomycin C on concentrations of vascular endothelial growth factor and its receptors in bladder cancer cells and in bladders of rats intravesically instilled with mitomycin C

AU - Verma, Avanti

AU - Degrado, Justin

AU - Hittelman, Adam B.

AU - Wheeler, Marcia A.

AU - Kaimakliotis, Hristos

AU - Weiss, Robert M.

PY - 2011/4

Y1 - 2011/4

N2 - OBJECTIVES • To examine, using in vitro and in vivo models, the largely unexamined effect of mitomycin C (MMC), an effective intravesical treatment for superficial bladder cancer and carcinoma in situ, on expression of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR-2), which mediates many of the angiogenic properties of VEGF. To measure, as a positive control, concentrations of the inhibitor of apoptosis, survivin, as an assessment of MMC effectiveness. To measure MMC-induced changes in proliferation in the presence and absence of VEGF-A small interfering RNA (siRNA). MATERIALS AND METHODS After treatment with increasing MMC concentrations (5-200 Âμg/mL), we measured proliferation, as well as VEGF, survivin, VEGF receptor-1 (VEGFR-1) and VEGFR-2 concentrations in RT-4 and T-24 bladder cancer cells. The effect of pre-treatment of VEGF siRNA and survivin siRNA on MMC-induced decreases in proliferation was measured. Urinary VEGF concentrations and bladder and kidney concentrations of VEGF-A, VEGFR-1, VEGFR-2 and interleukin-6 (IL-6) mRNA were measured in rats intravesically instilled with saline or MMC (200 Âμg/mL). RESULTS Although MMC treatment inhibited cell proliferation and decreased survivin mRNA expression in T-24 and RT4 cells, MMC (12-50 Âμg/mL) increased VEGF-A mRNA and VEGFR-2 mRNA and protein expression. Pre-treatment with VEGF-A siRNA or survivin siRNA before MMC treatment reduced proliferation more than MMC alone. MMC-induced reductions in proliferation were reduced additively by pre-treatment with survivin siRNA, but were potentiated by pre-treatment with VEGF-A siRNA. VEGFR-2 mRNA and protein concentrations and urinary VEGF concentrations were increased in bladders of rats instilled with MMC. CONCLUSIONS Intravesically instilled MMC increases urinary VEGF and bladder VEGFR-2 protein and mRNA in rats. MMC increases VEGF mRNA and VEGFR-2 protein and mRNA concentrations in bladder cancer cells. Therefore, we speculate that MMC could increase the angiogenic potential of both cancer and normal cells. In cancer cells this effect is largest at lower MMC concentrations. Combining MMC with agents that reduce EGF concentrations could be of value in treatment of transitional cell carcinoma of the bladder (TCC).

AB - OBJECTIVES • To examine, using in vitro and in vivo models, the largely unexamined effect of mitomycin C (MMC), an effective intravesical treatment for superficial bladder cancer and carcinoma in situ, on expression of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR-2), which mediates many of the angiogenic properties of VEGF. To measure, as a positive control, concentrations of the inhibitor of apoptosis, survivin, as an assessment of MMC effectiveness. To measure MMC-induced changes in proliferation in the presence and absence of VEGF-A small interfering RNA (siRNA). MATERIALS AND METHODS After treatment with increasing MMC concentrations (5-200 Âμg/mL), we measured proliferation, as well as VEGF, survivin, VEGF receptor-1 (VEGFR-1) and VEGFR-2 concentrations in RT-4 and T-24 bladder cancer cells. The effect of pre-treatment of VEGF siRNA and survivin siRNA on MMC-induced decreases in proliferation was measured. Urinary VEGF concentrations and bladder and kidney concentrations of VEGF-A, VEGFR-1, VEGFR-2 and interleukin-6 (IL-6) mRNA were measured in rats intravesically instilled with saline or MMC (200 Âμg/mL). RESULTS Although MMC treatment inhibited cell proliferation and decreased survivin mRNA expression in T-24 and RT4 cells, MMC (12-50 Âμg/mL) increased VEGF-A mRNA and VEGFR-2 mRNA and protein expression. Pre-treatment with VEGF-A siRNA or survivin siRNA before MMC treatment reduced proliferation more than MMC alone. MMC-induced reductions in proliferation were reduced additively by pre-treatment with survivin siRNA, but were potentiated by pre-treatment with VEGF-A siRNA. VEGFR-2 mRNA and protein concentrations and urinary VEGF concentrations were increased in bladders of rats instilled with MMC. CONCLUSIONS Intravesically instilled MMC increases urinary VEGF and bladder VEGFR-2 protein and mRNA in rats. MMC increases VEGF mRNA and VEGFR-2 protein and mRNA concentrations in bladder cancer cells. Therefore, we speculate that MMC could increase the angiogenic potential of both cancer and normal cells. In cancer cells this effect is largest at lower MMC concentrations. Combining MMC with agents that reduce EGF concentrations could be of value in treatment of transitional cell carcinoma of the bladder (TCC).

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KW - intravesical instillation

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KW - vascular endothelial growth factor 2

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