Selective targeting of lectins and their macropinocytosis in urothelial tumours : translation from in vitro to ex vivo

Resnik, Nataša (author), Višnjar, Tanja (author), Smrkolj, Tomaž (author), Erdani-Kreft, Mateja (author), Romih, Rok (author), Zupančič, Daša (author)

Urinary bladder cancer can be treated by intravesical application of therapeutic agents, but the specific targeting of cancer urothelial cells and the endocytotic pathways of the agents are not known. During carcinogenesis, the superficial urothelial cells exhibit changes in sugar residues on the apical plasma membranes. This can be exploited for selective targeting from the luminal side of the bladder. Here we show that the plant lectins Jacalin (from Artocarpus integrifolia), ACA (from Amaranthus caudatus) and DSA (from Datura stramonium) selectively bind to the apical plasma membrane of low- (RT4) and high-grade (T24) cancer urothelial cells in vitro and urothelial tumours ex vivo. The amount of lectin binding was significantly different between RT4 and T24 cells. Endocytosis of lectins was observed only in cancer urothelial cells and not in normal urothelial cells. Transmission electron microscopy analysis showed macropinosomes, endosome-like vesicles and multivesicular bodies filled with lectins in RT4 and T24 cells and also in cells of urothelial tumours ex vivo. Endocytosis of Jacalin and ACA in cancer cells was decreased in vitro after addition of inhibitor of macropinocytosis 5-(N-ethyl-N-isopropyl) amiloride (EIPA) and increased after stimulation of macropinocytosis with epidermal growth factor (EGF). Clathrin, caveolin and flotillin did not colocalise with lectins. These results confirm that the predominant mechanism of lectin endocytosis in cancer urothelial cells is macropinocytosis. Therefore, we propose that lectins in combination with conjugated therapeutic agents are promising tools for improved intravesical therapy by targeting cancer cells.

• Language: English
• Keywords: cancer urothelial cells, endocytosis, glycosylation, lectins, endometriosis, non-invasive diagnosis, macropinocytosis, urinary bladder tumours
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: MF - Faculty of Medicine
• Publication status: Published
• Publication version: Version of Record
• Year: 2023
• Number of pages: Str. 435–452
• Numbering: Vol. 160, iss. 5
• PID: 20.500.12556/RUL-152547
• UDC: 616-006:576
• ISSN on article: 0948-6143
• DOI: 10.1007/s00418-023-02224-2
• COBISS.SI-ID: 161188867
• Publication date in RUL: 27.11.2023

Record is a part of a journal

• Title: Histochemistry and cell biology
• Publisher: Springer Nature
• ISSN: 0948-6143
• COBISS.SI-ID: 1401876

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: urotelijske celice raka, endocitoza, glikozilacija, lektini, makropinocitoza, tumorji sečnega mehurja