Evans blue‑mediated white‑light detection of non‑muscle‑invasive bladder cancer: A preclinical feasibility and safety study using a rat bladder urothelial cell carcinoma model

Elsen,Sanne; Lerut,Evelyne; Van Der Aa,Frank; Van Cleynenbreugel,Ben; Van Poppel,Hendrik; De Witte,Peter

doi:10.3892/mco.2016.1043

Evans blue‑mediated white‑light detection of non‑muscle‑invasive bladder cancer: A preclinical feasibility and safety study using a rat bladder urothelial cell carcinoma model

Authors:
- Sanne Elsen
- Evelyne Lerut
- Frank Van Der Aa
- Ben Van Cleynenbreugel
- Hendrik Van Poppel
- Peter De Witte
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Affiliations: Laboratory of Molecular Biodiscovery, Faculty of Pharmaceutical Sciences, KU Leuven, B‑3000 Leuven, Belgium, Laboratory of Translational Cell and Tissue Research, Faculty of Medicine, KU Leuven, B‑3000 Leuven, Belgium, Department of Urology, University Hospital Leuven, B‑3000 Leuven, Belgium
Published online on: October 4, 2016 https://doi.org/10.3892/mco.2016.1043
Pages: 678-688
Copyright: © Elsen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Photodynamic diagnosis (PDD) improves the detection of non‑muscle‑invasive bladder cancer (NMIBC). However, white‑light (WL) cystoscopy remains the technique routinely used in urological clinics. A more cost‑effective but equally performant alternative to PDD may encompass the use of an intense tumoritropic dye in combination with WL cystoscopy. Using a preclinical setting, we investigated the practical aspects of the use of Evans blue (EB) dye for the possible future detection of NMIBC using WL cystoscopy. A solution of 1 and 5 mM EB was instilled into healthy and AY‑27 tumor‑bearing rat bladders. The bladders were then rapidly dissected and the inner walls were inspected for EB using WL stereomicroscopy. EB present in the bladders and the plasma was also quantified using high performance liquid chromatography. To assess the effects of repeated instillations on normal rat bladders, EB was instilled for 7 consecutive days, after which time the bladder wall was investigated histologically. To gain insight into the mechanisms underlying the selective accumulation of EB in malignant urothelium, RNA sequencing of urothelial tissue and subsequent comparative analysis were performed, with a specific focus on cell adhesion. The concentrations of EB were substantially higher in malignant bladders compared with those in healthy bladders, matching the blue staining of the inner bladder wall observed by stereomicroscopy. EB was equally present in the plasma of healthy and tumor‑bearing subjects, although at low concentrations. Importantly, EB did not cause any abnormalities in the urothelium after 7 days of repeated instillation in normal rats. RNA sequencing of the urothelium indicated an abnormal expression of several genes related to cell adhesion in malignant urothelium compared with the normal urothelium. Our findings may be important for future clinical developments in the field of diagnostics for bladder cancer. Implementing the more cost‑effective protocol of EB instillations in combination with WL cystoscopy may offer a benefit to patients as well as the healthcare system.

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