20.500.12556/DiRROS-15568 Evaluation of a novel plasmid for simultaneous gene electrotransfer-mediated silencing of CD105 and CD146 in combination with irradiation 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. CD105 CD146 plasmid gene electrotransfer CD105 CD146 plazmidi elektroprenos genov true false true MDPI Angleški jezik Slovenski jezik by Authors Neznano 2022-09-21 03:35:21 2022-09-21 14:35:43 2022-09-30 15:05:25 0000-00-00 00:00:00 2021 0 0 Opis vira z dne 19. 3. 2021; Nasl. z nasl. zaslona; str. 3069-1-3069-18 Vol. 22 2021 0000-00-00 Zaloznikova Objavljeno apc 0000-00-00 0000-00-00 2021-03-17 602.6/.7 1422-0067 10.3390/ijms22063069 56427011 https://www.mdpi.com/journal/ijms 1 e3535adb-393c-11ed-bea5-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=20591 RAZ_Savarin_Monika_2021.pdf RAZ_Savarin_Monika_2021.pdf 1 1AA6AAB5410529ADAF159488E9F205DE 4934f2a988b71fd3af68ebfc855b514a7f5fce56d5b4b7a76759d59190f78fc9 423ade63-3999-11ed-bea5-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=20605 Onkološki inštitut Ljubljana 0 0 0