Abstract
Intravesical mitomycin C (MMC) is commonly used to treat bladder cancer but is associated with local adverse effects. Here, we investigate the effects of MMC on release of urothelial mediators and production of inflammatory cytokines. Recovery and the effects of repeat treatment were also investigated. Urothelial cells were treated with MMC for 2 h at 37 °C. Immediately, 24 h and 7 days following MMC treatment, effects were assessed in terms of changes in ATP, acetylcholine and PGE2 release, and the presence of inflammatory cytokines and nitric oxide in incubation medium. Endpoints were also assessed 7 days after repeat MMC treatment. Immediately following MMC treatment at the clinical concentration (2 mg/mL), stretch-induced ATP release was significantly decreased, basal acetylcholine release was enhanced, while basal PGE2 was depressed and response to stretch increased. Twenty-four hours after treatment, basal acetylcholine was significantly increased ([MMC] ≥0.0002 mg/mL) and basal PGE2 enhanced (0.02 mg/mL). A 326-fold increase in interleukin-8 secretion and significant increase in nitric oxide release were also detected at 24 h. One week after single and repeat MMC treatment, urothelial ATP, acetylcholine and PGE2 had recovered while the increase in nitric oxide and interleukin-8 was still evident. These results indicate that urothelial mediator release is affected by MMC treatment with full recovery by 1 week. However, a concurrent increase in secretion of the inflammatory cytokine interleukin-8 and nitric oxide was also detected 24 h following treatment, which were still evident after the recovery period. These changes may play a role in the local adverse effects associated with intravesical MMC treatment.
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Abou El Hassan MA, Verheul HM, Jorna AS, Schalkwijk C, van Bezu J, van der Vijgh WJ, Bast A (2003) The new cardioprotector monohydroxyethylrutoside protects against doxorubicin-induced inflammatory effects in vitro. Br J Cancer 89:357–362. doi:10.1038/sj.bjc.6601022
Aizawa N, Igawa Y, Nishizawa O, Wyndaele JJ (2010) Effects of CL316,243, a beta 3-adrenoceptor agonist, and intravesical prostaglandin E2 on the primary bladder afferent activity of the rat. Neurourol Urodyn 29:771–776. doi:10.1002/nau.20826
Aizawa N, Igawa Y, Nishizawa O, Wyndaele JJ (2011) Effects of nitric oxide on the primary bladder afferent activities of the rat with and without intravesical acrolein treatment. Eur Urol 59:264–271. doi:10.1016/j.eururo.2010.10.035
Andersson MC, Tobin G, Giglio D (2008) Cholinergic nitric oxide release from the urinary bladder mucosa in cyclophosphamide-induced cystitis of the anaesthetized rat. Br J Pharmacol 153:1438–1444. doi:10.1038/bjp.2008.6
Aoki K, Hirayama A, Tanaka N, Yoneda T, Yoshida K, Fujimoto K, Hirao Y (2009) A higher level of prostaglandin E2 in the urinary bladder in young boys and boys with lower urinary tract obstruction. Biomed Res 30:343–347
Apodaca G, Balestreire E, Birder LA (2007) The uroepithelial-associated sensory web. Kidney Int 72:1057–1064. doi:10.1038/sj.ki.5002439
Aronsson P, Vesela R, Johnsson M, Tayem Y, Wsol V, Winder M, Tobin G (2014) Inhibition of nitric oxide synthase prevents muscarinic and purinergic functional changes and development of cyclophosphamide-induced cystitis in the rat. BioMed Res Int 2014:359179. doi:10.1155/2014/359179
Birder LA (2005) More than just a barrier: urothelium as a drug target for urinary bladder pain. Am J Physiol Ren Physiol 289:F489–F495. doi:10.1152/ajprenal.00467.2004
Birder LA (2010) Urothelial signaling. Auton Neurosci 153:33–40. doi:10.1016/j.autneu.2009.07.005
Birder LA, Apodaca G, De Groat WC, Kanai AJ (1998) Adrenergic- and capsaicin-evoked nitric oxide release from urothelium and afferent nerves in urinary bladder. Am J Physiol 275:F226–F229
Birder LA, Kanai AJ, Cruz F, Moore K, Fry CH (2010) Is the urothelium intelligent? Neurourol Urodyn 29:598–602. doi:10.1002/nau.20914
Burnstock G (2011) Therapeutic potential of purinergic signalling for diseases of the urinary tract. BJU Int 107:192–204. doi:10.1111/j.1464-410X.2010.09926.x
Bustamante S, Orensanz LM, Recio P, Carballido J, Garcia-Sacristan A, Prieto D, Hernandez M (2010) Functional evidence of nitrergic neurotransmission in the human urinary bladder neck. Neurosci Lett 477:91–94. doi:10.1016/j.neulet.2010.04.040
Cheng Y, Mansfield KJ, Sandow SL, Sadananda P, Burcher E, Moore KH (2011) Porcine bladder urothelial, myofibroblast, and detrusor muscle cells: characterization and ATP release. Front Pharmacol 2:27. doi:10.3389/fphar.2011.00027
Chou SF, Chang SW, Chuang JL (2007) Mitomycin C upregulates IL-8 and MCP-1 chemokine expression via mitogen-activated protein kinases in corneal fibroblasts. Invest Ophth Vis Sci 48:2009–2016. doi:10.1167/Iovs.06-0835
Cockayne DA et al (2000) Urinary bladder hyporeflexia and reduced pain-related behaviour in P2X3-deficient mice. Nature 407:1011–1015. doi:10.1038/35039519
Colombo R, Rocchini L, Suardi N, Benigni F, Colciago G, Bettiga A, Pellucchi F, Maccagnano C, Briganti A, Salonia A, Rigatti P, Montorsi F (2012) Neoadjuvant short-term intensive intravesical mitomycin C regimen compared with weekly schedule for low-grade recurrent non-muscle-invasive bladder cancer: preliminary results of a randomised phase 2 study. Eur Urol 62:797–802. doi:10.1016/j.eururo.2012.05.032
Dalton JT, Wientjes MG, Badalament RA, Drago JR, Au JL (1991) Pharmacokinetics of intravesical mitomycin C in superficial bladder cancer patients. Cancer Res 51:5144–5152
de Boer EC, Somogyi L, de Ruiter GJ, de Reijke TM, Kurth KH, Schamhart DH (1997) Role of interleukin-8 in onset of the immune response in intravesical BCG therapy for superficial bladder cancer. Urol Res 25:31–34
de Jongh R, van Koeveringe GA, van Kerrebroeck PE, Markerink-van Ittersum M, de Vente J, Gillespie JI (2007) The effects of exogenous prostaglandins and the identification of constitutive cyclooxygenase I and II immunoreactivity in the normal guinea pig bladder. BJU Int 100:419–429. doi:10.1111/j.1464-410X.2007.07011.x
Dokita S, Morgan WR, Wheeler MA, Yoshida M, Latifpour J, Weiss RM (1991) NG-nitro-L-arginine inhibits non-adrenergic, non-cholinergic relaxation in rabbit urethral smooth muscle. Life Sci 48:2429–2436
Duque JL, Loughlin KR (2000) An overview of the treatment of superficial bladder cancer. Intravesical chemotherapy. Urol Clin N Am 27:125–135
Ehren I, Hosseini A, Herulf M, Lundberg JO, Wiklund NP (1999) Measurement of luminal nitric oxide in bladder inflammation using a silicon balloon catheter: a novel minimally invasive method. Urology 54:264–267
Elmamoun MH, Christmas TJ, Woodhouse CR (2014) Destruction of the bladder by single dose mitomycin C for low-stage transitional cell carcinoma (TCC)—avoidance, recognition, management and consent. BJU Int 113:E34–E38. doi:10.1111/bju.12340
Giglio D, Ryberg AT, To K, Delbro DS, Tobin G (2005) Altered muscarinic receptor subtype expression and functional responses in cyclophosphamide induced cystitis in rats. Auton Neurosci 122:9–20. doi:10.1016/j.autneu.2005.07.005
Gillespie JI, Drake MJ (2004) The actions of sodium nitroprusside and the phosphodiesterase inhibitor dipyridamole on phasic activity in the isolated guinea-pig bladder. BJU Int 93:851–858. doi:10.1111/j.1464-410X.2003.04727.x
Hanna-Mitchell AT, Beckel JM, Barbadora S, Kanai AJ, de Groat WC, Birder LA (2007) Non-neuronal acetylcholine and urinary bladder urothelium. Life Sci 80:2298–2302. doi:10.1016/j.lfs.2007.02.010
Hayashi Y et al (2009) Bladder hyperactivity and increased excitability of bladder afferent neurons associated with reduced expression of Kv1.4 alpha-subunit in rats with cystitis. Am J Physiol Regul Int Comp Physiol 296:R1661–R1670. doi:10.1152/ajpregu.91054.2008
Heinrich M, Oberbach A, Schlichting N, Stolzenburg JU, Neuhaus J (2011) Cytokine effects on gap junction communication and connexin expression in human bladder smooth muscle cells and suburothelial myofibroblasts. PLoS One 6:e20792. doi:10.1371/journal.pone.0020792
Kamat AM, Lamm DL (2000) Intravesical therapy for bladder cancer. Urology 55:161–168
Kanai A et al (2007) Origin of spontaneous activity in neonatal and adult rat bladders and its enhancement by stretch and muscarinic agonists. Am J Physiol Ren Physiol 292:F1065–F1072. doi:10.1152/ajprenal.00229.2006
Kang SH, Chess-Williams R, Anoopkumar-Dukie S, McDermott C (2013) Induction of inflammatory cytokines and alteration of urothelial ATP, acetylcholine and prostaglandin E2 release by doxorubicin. Eur J Pharmacol 700:102–109. doi:10.1016/j.ejphar.2012.11.053
Koskela LR, Poljakovic M, Ehren I, Wiklund NP, de Verdier PJ (2012) Localization and expression of inducible nitric oxide synthase in patients after BCG treatment for bladder cancer. Nitric Oxide Biol Chem Off J Nitric Oxide Soc 27:185–191. doi:10.1016/j.niox.2012.07.001
Koya MP, Simon MA, Soloway MS (2006) Complications of intravesical therapy for urothelial cancer of the bladder. J Urol 175:2004–2010. doi:10.1016/S0022-5347(06)00264-3
Kullmann FA, Artim D, Beckel J, Barrick S, de Groat WC, Birder LA (2008) Heterogeneity of muscarinic receptor-mediated Ca2+ responses in cultured urothelial cells from rat. Am J Physiol Ren Physiol 294:F971–F981. doi:10.1152/ajprenal.00313.2007
Kumar V, Chapple CR, Rosario D, Tophill PR, Chess-Williams R (2010) In vitro release of adenosine triphosphate from the urothelium of human bladders with detrusor overactivity, both neurogenic and idiopathic. Eur Urol 57:1087–1092. doi:10.1016/j.eururo.2009.11.042
Kumar V, Chapple CR, Surprenant AM, Chess-Williams R (2007) Enhanced adenosine triphosphate release from the urothelium of patients with painful bladder syndrome: a possible pathophysiological explanation. J Urol 178:1533–1536. doi:10.1016/j.juro.2007.05.116
Logadottir Y, Delbro D, Fall M, Gjertsson I, Jirholt P, Lindholm C, Peeker R (2014) Cytokine expression in patients with bladder pain syndrome/interstitial cystitis ESSIC type 3C. J Urol 192:1564–1568. doi:10.1016/j.juro.2014.04.099
Logadottir YR, Ehren I, Fall M, Wiklund NP, Peeker R, Hanno PM (2004) Intravesical nitric oxide production discriminates between classic and nonulcer interstitial cystitis. J Urol 171:1148–1150. doi:10.1097/01.ju.0000110501.96416.40, discussion 1150-1141
Logan C, Brown M, Hayne D (2012) Intravesical therapies for bladder cancer—indications and limitations. BJU Int 110(Suppl 4):12–21. doi:10.1111/j.1464-410X.2012.11619.x
Lundberg JO et al (1996) Elevated nitric oxide in the urinary bladder in infectious and noninfectious cystitis. Urology 48:700–702. doi:10.1016/S0090-4295(96)00423-2
Maggi CA et al (1988) Prostanoids modulate reflex micturition by acting through capsaicin-sensitive afferents. Eur J Pharmacol 145:105–112
Mansfield KJ, Hughes JR (2014a) Effect of inflammatory mediators on ATP release of human urothelial RT4 cells. BioMed Res Int 2014:182862. doi:10.1155/2014/182862
Mansfield KJ, Hughes JR (2014b) P2Y receptor modulation of ATP release in the urothelium. BioMed Res Int 2014:830374. doi:10.1155/2014/830374
McDermott C, Chess-Williams R, Grant GD, Perkins AV, McFarland AJ, Davey AK, Anoopkumar-Dukie S (2012) Effects of Pseudomonas aeruginosa virulence factor pyocyanin on human urothelial cell function and viability. J Urol 187:1087–1093. doi:10.1016/j.juro.2011.10.129
Moon A (2002) Influence of nitric oxide signalling pathways on pre-contracted human detrusor smooth muscle in vitro. BJU Int 89:942–949
Moro C, Leeds C, Chess-Williams R (2012) Contractile activity of the bladder urothelium/lamina propria and its regulation by nitric oxide. Eur J Pharmacol 674:445–449. doi:10.1016/j.ejphar.2011.11.020
Nomiya M et al (2012) Increased bladder activity is associated with elevated oxidative stress markers and proinflammatory cytokines in a rat model of atherosclerosis-induced chronic bladder ischemia. Neurourol Urodyn 31:185–189. doi:10.1002/nau.21191
Pouya M, Opsomer RJ, Wese FX, Saleh M, Donnay M, Van Cangh PJ (1996) Complications of intravesical mitomycin chemotherapy, apropos of 2 clinical case reports. Acta Urol Belg 64:47–53
Souslova V et al (2000) Warm-coding deficits and aberrant inflammatory pain in mice lacking P2X3 receptors. Nature 407:1015–1017. doi:10.1038/35039526
Tanaka I, Nagase K, Tanase K, Aoki Y, Akino H, Yokoyama O (2011) Modulation of stretch evoked adenosine triphosphate release from bladder epithelium by prostaglandin E(2). J Urol 185:341–346. doi:10.1016/j.juro.2010.09.042
Thrasher JB, Crawford ED (1992) Complications of intravesical chemotherapy. Urol Clin N Am 19:529–539
Wientjes MG, Badalament RA, Wang RC, Hassan F, Au JL (1993) Penetration of mitomycin C in human bladder. Cancer Res 53:3314–3320
Wientjes MG, Dalton JT, Badalament RA, Drago JR, Au JL (1991) Bladder wall penetration of intravesical mitomycin C in dogs. Cancer Res 51:4347–4354
Wyndaele JJ, De Wachter S (2003) The basics behind bladder pain: a review of data on lower urinary tract sensations. Int J Urol Off J Jpn Urol Assoc 10(Suppl):S49–S55
Yokoyama O (2010) Pharmacological and genetic analysis of mechanisms underlying detrusor overactivity in rats. Neurourol Urodyn 29:107–111. doi:10.1002/nau.20746
Yoshimura N, Seki S, Chancellor MB, de Groat WC, Ueda T (2002) Targeting afferent hyperexcitability for therapy of the painful bladder syndrome. Urology 59:61–67
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This study was funded by a project grant from Cancer Council Queensland and the National Health and Medical Research Council of Australia (527502).
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Kang, S.H., Chess-Williams, R., Anoopkumar-Dukie, S. et al. Recovery of urothelial mediator release but prolonged elevations in interleukin-8 and nitric oxide secretion following mitomycin C treatment. Naunyn-Schmiedeberg's Arch Pharmacol 388, 781–791 (2015). https://doi.org/10.1007/s00210-015-1092-7
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DOI: https://doi.org/10.1007/s00210-015-1092-7