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Article

Evaluation of a Novel Plasmid for Simultaneous Gene Electrotransfer-Mediated Silencing of CD105 and CD146 in Combination with Irradiation

1
Department of Experimental Oncology, Institute of Oncology Ljubljana, 1000 Ljubljana, Slovenia
2
Biotechnical Faculty, University of Ljubljana, 1000 Ljubljana, Slovenia
3
Faculty of Health Sciences, University of Ljubljana, 1000 Ljubljana, Slovenia
4
Faculty of Health Sciences, University of Primorska, 6310 Izola, Slovenia
*
Authors to whom correspondence should be addressed.
Academic Editor: Lukas J.A.C. Hawinkels
Int. J. Mol. Sci. 2021, 22(6), 3069; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22063069
Received: 1 February 2021 / Revised: 12 March 2021 / Accepted: 13 March 2021 / Published: 17 March 2021
(This article belongs to the Special Issue Endoglin in Health and Disease)
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. View Full-Text
Keywords: CD105; CD146; plasmid; gene electrotransfer; antibiotic-free; irradiation CD105; CD146; plasmid; gene electrotransfer; antibiotic-free; irradiation
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MDPI and ACS Style

Savarin, M.; Kamensek, U.; Znidar, K.; Todorovic, V.; Sersa, G.; Cemazar, M. Evaluation of a Novel Plasmid for Simultaneous Gene Electrotransfer-Mediated Silencing of CD105 and CD146 in Combination with Irradiation. Int. J. Mol. Sci. 2021, 22, 3069. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22063069

AMA Style

Savarin M, Kamensek U, Znidar K, Todorovic V, Sersa G, Cemazar M. Evaluation of a Novel Plasmid for Simultaneous Gene Electrotransfer-Mediated Silencing of CD105 and CD146 in Combination with Irradiation. International Journal of Molecular Sciences. 2021; 22(6):3069. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22063069

Chicago/Turabian Style

Savarin, Monika; Kamensek, Urska; Znidar, Katarina; Todorovic, Vesna; Sersa, Gregor; Cemazar, Maja. 2021. "Evaluation of a Novel Plasmid for Simultaneous Gene Electrotransfer-Mediated Silencing of CD105 and CD146 in Combination with Irradiation" Int. J. Mol. Sci. 22, no. 6: 3069. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22063069

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