Heterogeneity of uroplakin localization in human normal urothelium, papilloma and papillary carcinoma

Abstract

BackgroundUroplakins are differentiation-related membrane proteins of urothelium. We compared uroplakin expression and ultrastructural localization in human normal urothelium, papilloma and papillary carcinoma. Because of high recurrence rate of these tumours, treated by transurethral resection, we investigated urothelial tumour, resection border and uninvolved urothelium.Patients and methodsUrinary bladder samples were obtained from tumour free control subjects and patients with papilloma and papillary carcinoma. Immunohistochemical and immunoelectron labelling of uroplakins were performed.ResultsIn normal human urothelium with continuous uroplakin-positive superficial cell layer uroplakins were localized to flattened mature fusiform vesicles and apical plasma membrane of umbrella cells. Diverse uroplakin expression was found in papilloma and papillary carcinoma. Three aberrant differentiation stages of urothelial cells, not found in normal urothelium, were recognized in tumours. Diverse uroplakin expression and aberrant differentiation were occasionally found in resection border and in uninvolved urothelium.ConclusionsWe demonstrated here that uroplakin expression and localization in urothelial tumours is altered when compared to normal urothelium. In patients with papilloma and papillary carcinoma immunolabelling of uroplakins at ultrastructural level shows aberrant urothelial differentiation. It is possible that aberrant differentiation stages of urothelial cells in resection border and in uninvolved urothelium contribute to high recurrence rate.

Correlation between Urothelial Differentiation and Sensory Proteins P2X3, P2X5, TRPV1, and TRPV4 in Normal Urothelium and Papillary Carcinoma of Human Bladder

“…Apical surface was scalloped, with microridges covering the umbrella cells. These characteristics demonstrate terminal differentiation of the urothelium, as proposed previously [21]. In low grade carcinoma decrease in uroplakin expression was detected and altered urothelial apical surface appearance was observed.…”

Highly Selective Anti-Cancer Activity of Cholesterol-Interacting Agents Methyl-β-Cyclodextrin and Ostreolysin A/Pleurotolysin B Protein Complex on Urothelial Cancer Cells

“…Thus, two human urothelial cell lines with different levels of cancer transformation, as T24 and RT4 cells, were analyzed for cytotoxicity along with NPU cells in the present study. Since normal human urothelium is difficult-to-obtain tissue, we used normal porcine urothelial cell culture, which shows identical differentiation markers as well as cell biological and histological similarities to human urothelium [ 35 , 50 ].…”

“…To verify the influence of potential interspecies variability of cholesterol content, we measured the cholesterol content also in invasive and noninvasive mouse urothelial cell lines. We compared the sensitivity to MCD and OlyA/PlyB of T24 human urothelial cancer cells (as a high-grade invasive urothelial carcinoma model), RT4 human urothelial cancer cells (as a low-grade noninvasive papillary carcinoma model), and NPU cells, which morphologically and physiologically closely resemble normal human urothelium [ 33 , 34 , 35 ]. We took advantage of the unique mechanism of OlyA/PlyB protein complex to allow, on the one hand, efficient immunolabeling of cholesterol/ sphingomyelin-enriched membrane domains [ 3 , 32 , 36 ], and on the other hand, controlled pore-formation in these domains [ 28 , 29 , 32 ].…”

Co‐culturing porcine normal urothelial cells, urinary bladder fibroblasts and smooth muscle cells for tissue engineering research

“…The immunofluorescence reaction was performed as described previously (Kreft et al, 2005b). Each antibody used in this study has been previously extensively tested in our lab by performing immunofluorescence as well as immunohistochemistry by biotinylated secondary antibodies on cell cultures and on cryo‐ and paraffin‐sections of pig, mouse, rat, and human urinary bladder urothelium tissue sections (unpublished research and Kreft et al, , , , , 2010; Visnjar et al, ; Visnjar and Kreft, , ; Zupancic et al, , , ; Zupancic and Romih, ). The panel of antibodies we used was: CK 7 (mouse monoclonal antibody, diluted 1:20, Dako, Glostrup, Denmark), vimentin (rabbit polyclonal antibody, diluted 1:20, Dako), desmin (rabbit polyclonal antibody, diluted 1:40, Sigma), collagen I (mouse monoclonal antibody, diluted 1:200, Sigma), occludin (rabbit polyclonal antibody, diluted 1:20, Zymed Laboratories, San Francisco, CA, USA), E‐cadherin (mouse monoclonal antibody, diluted 1:20, Transduction Laboratories, Lexington, KY, USA), uroplakins (rabbit polyclonal antibody, diluted 1:10.000, a kind gift from Prof. dr. T.T.…”