Cytotoxic activity of LLO Y406A is targeted to the plasma membrane of cancer urothelial cells

Resnik, Nataša (author), Tratnjek, Larisa (author), Erdani-Kreft, Mateja (author), Kisovec, Matic (author), Aden, Saša (author), Bedina Zavec, Apolonija (author), Anderluh, Gregor (author), Podobnik, Marjetka (author), Veranič, Peter (author)

Identification of novel agents for bladder cancer treatment is highly desirable due to the high incidence of tumor recurrence and the risk of progression to muscle-invasive disease. The key feature of the cholesterol-dependent toxin listeriolysin O mutant (LLO Y406A) is its preferential activity at pH 5.7, which could be exploited either directly for selective targeting of cancer cells or the release of accumulated therapeutics from acidic endosomes. Therefore, our goal was to compare the cytotoxic effect of LLO Y406A on cancer cells (RT4) and normal urothelial cells (NPU), and to identify which cell membranes are the primary target of LLO Y406A by viability assays, life-cell imaging, fluorescence, and electron microscopy. LLO Y406A decreased viability, altered cell morphology, provoked membrane blebbing, and induced apoptosis in RT4 cells, while it did not affect NPU cells. LLO Y406A did not cause endosomal escape in RT4 cells, while the plasma membrane of RT4 cells was revealed as the primary target of LLO Y406A. It has been concluded that LLO Y406A has the ability to selectively eliminate cancer urothelial cells through pore-forming activity at the plasma membrane, without cytotoxic effects on normal urothelial cells. This promising selective activity merits further testing as an anti-cancer agent.

• Language: English
• Keywords: bladder cancer, cancer urothelial cells, normal urothelial cells, listeriolysin O, blebbing, calcium influx, pore-forming toxin, cytolysin, cytotoxicity
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: MF - Faculty of Medicine
• Publication status: Published
• Publication version: Version of Record
• Year: 2021
• Number of pages: 15 str.
• Numbering: Vol. 22, iss. 7, art. 3305
• PID: 20.500.12556/RUL-135019
• UDC: 577
• ISSN on article: 1422-0067
• DOI: 10.3390/ijms22073305
• COBISS.SI-ID: 57572867
• Publication date in RUL: 17.02.2022

Record is a part of a journal

• Title: International journal of molecular sciences
• Shortened title: Int. j. mol. sci.
• Publisher: MDPI
• ISSN: 1422-0067
• COBISS.SI-ID: 2779162

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.
• Licensing start date: 01.04.2021

Secondary language

• Language: Slovenian
• Keywords: biokemija, rakave celice, listeriolizin O, toksini