Gemcitabine: Selective cytotoxicity, induction of inflammation and effects on urothelial function.

Intravesical gemcitabine has recently been introduced for the treatment of superficial bladder cancer and has a favourable efficacy and toxicity profile in comparison to mitomycin c (MMC), the most commonly used chemotherapeutic agent. The aim of this study was to assess the cytotoxic potency of gemcitabine in comparison to MMC in urothelial cell lines derived from non-malignant (UROtsa) and malignant (RT4 and T24) tissues to assess selectivity. Cells were treated with gemcitabine or mitomycin C at concentrations up to the clinical doses for 1 or 2h respectively (clinical duration). Treatment combined with hyperthermia was also examined. Cell viability, ROS formation, urothelial function (ATP, acetylcholine and PGE2 release) and secretion of inflammatory cytokines were assessed. Gemcitabine displayed a high cytotoxic selectivity for the two malignant cell lines (RT4, T24) compared to the non-malignant urothelial cells (UROtsa, proliferative and non-proliferative). In contrast, the cytotoxic effects of MMC were non-selective with equivalent potency in each of the cell lines. The cytotoxic effect of gemcitabine in the malignant cell lines was associated with an elevation in free radical formation and was significantly decreased in the presence of an equilibrative nucleoside transporter inhibitor. Transient changes in urothelial ATP and PGE2 release were observed, with significant increase in release of interleukin-6, interleukin-8 and interleukin-1ß from urothelial cells treated with gemcitabine. The selectivity of gemcitabine for malignant urothelial cells may account for the less frequent adverse urological effects with comparison to other commonly used chemotherapeutic agents.

Enhanced urothelial ATP release and contraction following intravesical treatment with the cytotoxic drug, doxorubicin.

Intravesical administration of the cytotoxic drug doxorubicin is a common treatment for superficial carcinoma of the bladder, but it is associated with significant urological adverse effects. The aim of this study was to identify doxorubicin-induced changes in the local mechanisms involved in regulating bladder function. As a model of intravesical doxorubicin administration in patients, doxorubicin (1 mg/mL) was applied to the luminal surface of porcine bladders for 60 min. Following treatment, the release of urothelial/lamina propria mediators (acetylcholine (Ach), ATP and prostaglandin E2 (PGE2) and contractile responses of isolated tissue strips was investigated. Doxorubicin pretreatment did not affect contractile responses of detrusor muscle to carbachol, but did enhance neurogenic detrusor responses to electrical field stimulation (219 % at 5 Hz). Contractions of isolated strips of urothelium/lamina propria to carbachol were also enhanced (30 %) in tissues from doxorubicin pretreated bladders. Isolated strips of urothelium/lamina propria from control bladders demonstrated a basal release of all three mediators (Ach > ATP > PGE2), with increased release of ATP when tissues were stretched. In tissues from doxorubicin-pretreated bladders, the basal release of ATP was significantly enhanced (sevenfold), while the release of acetylcholine and PGE2 was not affected. The application of luminal doxorubicin, under conditions that mimic intravesical administration to patients, affects urothelial/lamina propria function (increased contractile activity and ATP release) and enhances efferent neurotransmission without affecting detrusor smooth muscle. These actions would enhance bladder contractile activity and sensory nerve activity and may explain the adverse urological effects observed in patients following intravesical doxorubicin treatment.