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Title:Evaluation of a novel plasmid for simultaneous gene electrotransfer-mediated silencing of CD105 and CD146 in combination with irradiation
Authors:ID Savarin, Monika (Author)
ID Kamenšek, Urška (Author)
ID Žnidar, Katarina (Author)
ID Todorović, Vesna (Author)
ID Serša, Gregor (Author)
ID Čemažar, Maja (Author)
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MD5: 1AA6AAB5410529ADAF159488E9F205DE
Typology:1.01 - Original Scientific Article
Organization:Logo OI - Institute of Oncology
Abstract:Targeting tumor vasculature through specific endothelial cell markers represents a promising approach for cancer treatment. Here our aim was to construct an antibiotic resistance gene-free plasmid encoding shRNAs to simultaneously target two endothelial cell markers, CD105 and CD146, and to test its functionality and therapeutic potential in vitro when delivered by gene electrotransfer (GET) and combined with irradiation (IR). Functionality of the plasmid was evaluated by determining the silencing of the targeted genes using qRT-PCR. Antiproliferative and antiangiogenic effects were determined by the cytotoxicity assay tube formation assay and wound healing assay in murine endothelial cells 2H-11. The functionality of the plasmid construct was also evaluated in malignant melanoma tumor cell line B16F10. Additionally, potential activation of immune response was measured by induction of DNA sensor STING and proinflammatory cytokines by qRT-PCR in endothelial cells 2H-11. We demonstrated that the plasmid construction was successful and can efficiently silence the expression of the two targeted genes. As a consequence of silencing, reduced migration rate and angiogenic potential was confirmed in 2H-11 endothelial cells. Furthermore, induction of DNA sensor STING and proinflammatory cytokines were determined, which could add to the therapeutic effectiveness when used in vivo. To conclude, we successfully constructed a novel plasmid DNA with two shRNAs, which holds a great promise for further in vivo testing.
Keywords:CD105, CD146, plasmid, gene electrotransfer
Publication status:Published
Publication version:Version of Record
Publication date:17.03.2021
Year of publishing:2021
Number of pages:str. 3069-1-3069-18
Numbering:Vol. 22
PID:20.500.12556/DiRROS-15568 New window
ISSN on article:1422-0067
DOI:10.3390/ijms22063069 New window
COBISS.SI-ID:56427011 New window
Copyright:by Authors
Note:Opis vira z dne 19. 3. 2021; Nasl. z nasl. zaslona;
Publication date in DiRROS:21.09.2022
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Record is a part of a journal

Title:International journal of molecular sciences
Shortened title:Int. j. mol. sci.
COBISS.SI-ID:2779162 New window

Document is financed by a project

Funder:ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
Project number:Z3-1873-2019
Name:Nov pristop žilno ciljane terapije, kjer z gensko terapijo utišamo dve neodvisni signalni poti,v kombinaciji z radioterapijo

Funder:ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
Project number:P3-0003-2015
Name:Razvoj in ovrednotenje novih terapij za zdravljenje malignih tumorjev


License:CC BY 4.0, Creative Commons Attribution 4.0 International
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:21.09.2022

Secondary language

Keywords:CD105, CD146, plazmidi, elektroprenos genov