Elsen Sanne, Lerut Evelyne, Van Der Aa Frank, Van Cleynenbreugel Ben, Van Poppel Hendrik, De Witte Peter
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.
Mol Clin Oncol. 2016 Dec;5(6):678-688. doi: 10.3892/mco.2016.1043. Epub 2016 Oct 4.
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.
光动力诊断（PDD）可提高非肌层浸润性膀胱癌（NMIBC）的检测率。然而，白光（WL）膀胱镜检查仍是泌尿外科诊所常规使用的技术。一种更具成本效益但性能相当的PDD替代方法可能包括使用一种强肿瘤亲和性染料与WL膀胱镜检查相结合。在临床前环境中，我们研究了使用伊文思蓝（EB）染料通过WL膀胱镜检查未来可能检测NMIBC的实际情况。将1 mM和5 mM的EB溶液注入健康大鼠和携带AY - 27肿瘤的大鼠膀胱中。然后迅速解剖膀胱，使用WL体视显微镜检查膀胱内壁是否有EB。还使用高效液相色谱法定量膀胱和血浆中存在的EB。为了评估重复灌注对正常大鼠膀胱的影响，连续7天灌注EB，之后对膀胱壁进行组织学研究。为了深入了解EB在恶性尿路上皮中选择性积累的潜在机制，对尿路上皮组织进行了RNA测序及后续比较分析，特别关注细胞黏附。与健康膀胱相比，恶性膀胱中EB的浓度显著更高，这与体视显微镜观察到的膀胱内壁蓝色染色相符。健康和患肿瘤大鼠的血浆中均有EB，尽管浓度较低。重要的是，在正常大鼠中重复灌注7天后，EB未引起尿路上皮的任何异常。尿路上皮的RNA测序表明，与正常尿路上皮相比，恶性尿路上皮中几个与细胞黏附相关的基因表达异常。我们的发现可能对膀胱癌诊断领域未来的临床发展具有重要意义。实施更具成本效益的EB灌注方案与WL膀胱镜检查相结合可能对患者和医疗系统都有益处。
