51. Effect of ionizing radiation on human skeletal muscle precursor cellsMihaela Jurdana, Maja Čemažar, Katarina Pegan, Tomaž Marš, 2013, published scientific conference contribution Abstract: Background. Long term effects of different doses of ionizing radiation on human skeletal muscle myoblast proliferation, cytokine signalling and stress response capacity were studied in primary cell cultures.Materials and methods. Human skeletal muscle myoblasts obtained from muscle biopsies were cultured and irradiated with a Darpac 2000 X-ray unit at doses of 4, 6 and 8 Gy. Acute effects of radiation were studied by interleukin - 6 (IL-6) release and stress response detected by the heat shock protein (HSP) level, while long term effects were followed by proliferation capacity and cell death.Results. Compared with non-irradiated control and cells treated with inhibitor of cell proliferation Ara C, myoblast proliferation decreased 72 h post-irradiation, this effect was more pronounced with increasing doses. Post-irradiation myoblast survival determined by measurement of released LDH enzyme activity revealed increased activity after exposure to irradiation. The acute response of myoblasts to lower doses of irradiation (4 and 6 Gy) was decreased secretion of constitutive IL-6. Higher doses of irradiation triggered a stress response in myoblasts, determined by increased levels of stress markers (HSPs 27 and 70).Conclusions. Our results show that myoblasts are sensitive to irradiation in terms of their proliferation capacity and capacity to secret IL-6. Since myoblast proliferation and differentiation are a key stage in muscle regeneration, this effect of irradiation needs to be taken in account, particularly in certain clinical conditions.
Keywords: myoblasts, irradiation, proliferation, interleukin 6, muscle regeneration, apoptosis
Published in RUP: 30.12.2015; Views: 3954; Downloads: 150
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52. Endoglin (CD105) silencing mediated by shRNA under the control of Endothelin-1 promoter for targeted gene therapy of MelanomaNataša Tešić, Urška Kamenšek, Gregor Serša, Simona Kranjc Brezar, Monika Savarin, Urša Lampreht Tratar, Véronique Préat, Gaëlle Vandermeulen, Miha Butinar, Boris Turk, Maja Čemažar, 2015, original scientific article Abstract: Endoglin (CD105), a transforming growth factor (TGF)-% coreceptor, and endothelin-1, a vasoconstrictor peptide, are both overexpressed in tumor endothelial and melanoma cells. Their targeting is therefore a promising therapeutic approach for melanoma tumors. The aim of our study was to construct a eukaryotic expression plasmid encoding the shRNA molecules against CD105 under the control of endothelin-1 promoter and to evaluate its therapeutic potential both in vitro in murine B16F10-luc melanoma and SVEC4-10 endothelial cells and in vivo in mice bearing highly metastatic B16F10-luc tumors. Plasmid encoding shRNA against CD105 under the control of the constitutive U6 promoter was used as a control. We demonstrated the antiproliferative and antiangiogenic effects of both plasmids in SVEC4-10 cells, as well as a moderate antitumor and pronounced antimetastatic effect in B16F10-luc tumors in vivo. Our results provide evidence that targeting melanoma with shRNA molecules against CD105 under the control of endothelin-1 promoter is a feasible and effective treatment, especially for the reduction of metastatic spread.
Keywords: melanoma, electroporation, endoglin (CD105), endothelin-1, gene therapy
Published in RUP: 14.10.2015; Views: 3793; Downloads: 131
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56. Biological properties of melanoma and endothelial cells after plasmid AMEP gene electrotransfer depend on integrin quantity on cellsMaša Bošnjak, Lara Prosen, Tanja Jesenko, Tanja Blagus, Boštjan Markelc, Maja Čemažar, Céline Bouquet, Gregor Serša, 2013, original scientific article Keywords: antiangiogenic, integrins, gene electrotransfer, human endothelial cells, murine melanoma cells, plasmid AMEP
Published in RUP: 15.10.2013; Views: 3965; Downloads: 117
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