Effects of pseudomonas aeruginosa virulence factor pyocyanin on human urothelial cell function and viability

Catherine McDermott, Russ Chess-Williams, Gary D. Grant, Anthony V. Perkins, Amelia J. McFarland, Andrew K. Davey, Shailendra Anoopkumar-Dukie

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Abstract

Purpose: We determined the effects of Pseudomonas aeruginosa virulence factor pyocyanin on human urothelial cell viability and function in vitro. Materials and Methods: RT4 urothelial cells were treated with pyocyanin (1 to 100 μM) for 24 hours. After exposure the treatment effects were measured according to certain end points, including changes in urothelial cell viability, reactive oxygen species formation, caspase-3 activity, basal and stimulated adenosine triphosphate release, SA-β-gal activity and detection of acidic vesicular organelles. Results: The 24-hour pyocyanin treatment resulted in a concentration dependent decrease in cell viability at concentrations of 25 μM or greater, and increases in reactive oxygen species formation and caspase-3 activity at 25 μM or greater. Basal adenosine triphosphate release was significantly decreased at all tested pyocyanin concentrations while stimulated adenosine triphosphate release was significantly inhibited at pyocyanin concentrations of 12.5 μM or greater with no significant stimulated release at 100 μM. Pyocyanin treated RT4 cells showed morphological characteristics associated with cellular senescence, including SA-β-gal expression. This effect was not evident at 100 μM pyocyanin and may have been due to apoptotic cell death, as indicated by increased caspase-3 activity. An increase in acridine orange stained vesicular-like organelles was observed in RT4 urothelial cells after pyocyanin treatment. Conclusions: Exposure to pyocyanin alters urothelial cell viability, reactive oxygen species production and caspase-3 activity. Treatment also results in cellular senescence, which may affect the ability of urothelium to repair during infection. The virulence factor depressed stimulated adenosine triphosphate release, which to our knowledge is a novel finding with implications for awareness of bladder filling in patients with P. aeruginosa urinary tract infection.

Original languageEnglish
Pages (from-to)1087-1093
Number of pages7
JournalJournal of Urology
Volume187
Issue number3
DOIs
Publication statusPublished - Mar 2012

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Pyocyanine
Virulence Factors
Pseudomonas aeruginosa
Cell Survival
Caspase 3
Adenosine Triphosphate
Reactive Oxygen Species
Cell Aging
Organelles
Urothelium
Acridine Orange
Therapeutics
Urinary Tract Infections
Urinary Bladder
Cell Death

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McDermott, Catherine ; Chess-Williams, Russ ; Grant, Gary D. ; Perkins, Anthony V. ; McFarland, Amelia J. ; Davey, Andrew K. ; Anoopkumar-Dukie, Shailendra. / Effects of pseudomonas aeruginosa virulence factor pyocyanin on human urothelial cell function and viability. In: Journal of Urology. 2012 ; Vol. 187, No. 3. pp. 1087-1093.
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abstract = "Purpose: We determined the effects of Pseudomonas aeruginosa virulence factor pyocyanin on human urothelial cell viability and function in vitro. Materials and Methods: RT4 urothelial cells were treated with pyocyanin (1 to 100 μM) for 24 hours. After exposure the treatment effects were measured according to certain end points, including changes in urothelial cell viability, reactive oxygen species formation, caspase-3 activity, basal and stimulated adenosine triphosphate release, SA-β-gal activity and detection of acidic vesicular organelles. Results: The 24-hour pyocyanin treatment resulted in a concentration dependent decrease in cell viability at concentrations of 25 μM or greater, and increases in reactive oxygen species formation and caspase-3 activity at 25 μM or greater. Basal adenosine triphosphate release was significantly decreased at all tested pyocyanin concentrations while stimulated adenosine triphosphate release was significantly inhibited at pyocyanin concentrations of 12.5 μM or greater with no significant stimulated release at 100 μM. Pyocyanin treated RT4 cells showed morphological characteristics associated with cellular senescence, including SA-β-gal expression. This effect was not evident at 100 μM pyocyanin and may have been due to apoptotic cell death, as indicated by increased caspase-3 activity. An increase in acridine orange stained vesicular-like organelles was observed in RT4 urothelial cells after pyocyanin treatment. Conclusions: Exposure to pyocyanin alters urothelial cell viability, reactive oxygen species production and caspase-3 activity. Treatment also results in cellular senescence, which may affect the ability of urothelium to repair during infection. The virulence factor depressed stimulated adenosine triphosphate release, which to our knowledge is a novel finding with implications for awareness of bladder filling in patients with P. aeruginosa urinary tract infection.",
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Effects of pseudomonas aeruginosa virulence factor pyocyanin on human urothelial cell function and viability. / McDermott, Catherine; Chess-Williams, Russ; Grant, Gary D.; Perkins, Anthony V.; McFarland, Amelia J.; Davey, Andrew K.; Anoopkumar-Dukie, Shailendra.

In: Journal of Urology, Vol. 187, No. 3, 03.2012, p. 1087-1093.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Chess-Williams, Russ

AU - Grant, Gary D.

AU - Perkins, Anthony V.

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AU - Davey, Andrew K.

AU - Anoopkumar-Dukie, Shailendra

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N2 - Purpose: We determined the effects of Pseudomonas aeruginosa virulence factor pyocyanin on human urothelial cell viability and function in vitro. Materials and Methods: RT4 urothelial cells were treated with pyocyanin (1 to 100 μM) for 24 hours. After exposure the treatment effects were measured according to certain end points, including changes in urothelial cell viability, reactive oxygen species formation, caspase-3 activity, basal and stimulated adenosine triphosphate release, SA-β-gal activity and detection of acidic vesicular organelles. Results: The 24-hour pyocyanin treatment resulted in a concentration dependent decrease in cell viability at concentrations of 25 μM or greater, and increases in reactive oxygen species formation and caspase-3 activity at 25 μM or greater. Basal adenosine triphosphate release was significantly decreased at all tested pyocyanin concentrations while stimulated adenosine triphosphate release was significantly inhibited at pyocyanin concentrations of 12.5 μM or greater with no significant stimulated release at 100 μM. Pyocyanin treated RT4 cells showed morphological characteristics associated with cellular senescence, including SA-β-gal expression. This effect was not evident at 100 μM pyocyanin and may have been due to apoptotic cell death, as indicated by increased caspase-3 activity. An increase in acridine orange stained vesicular-like organelles was observed in RT4 urothelial cells after pyocyanin treatment. Conclusions: Exposure to pyocyanin alters urothelial cell viability, reactive oxygen species production and caspase-3 activity. Treatment also results in cellular senescence, which may affect the ability of urothelium to repair during infection. The virulence factor depressed stimulated adenosine triphosphate release, which to our knowledge is a novel finding with implications for awareness of bladder filling in patients with P. aeruginosa urinary tract infection.

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