Esistance observed in exosomes treated cells is correlated for the activation on the PI3K/AKT survival

Esistance observed in exosomes treated cells is correlated for the activation on the PI3K/AKT survival signalling pathway that involves the FoxO1 phosphorylation. Intriguingly, the Western blot analysis with the microvesicles purified from SH-SY5Y culture medium shows the presence of activated AKT kinase, i.e. phosphorylated on each serine 473 and threonine 308 residues. Summary/Conclusion: These observations indicate that exosomes may perhaps induce radiation resistance in SH-SY5Y cells by mechanisms involving FoxO1 phosphorylation, hence blocking the apoptotic method triggered by radiation. Our hypothesis is the fact that this pathway is activated or reinforced by the uptake of exosomes carrying phosphorylated AKT. Funding: This study was funded by Italian Ministry of Foreign Affairs and international Cooperation (grant: PGR00782).PS08.Extracellular vesicles shedding in response to chemotherapy in melanoma promotes tumour growth immediately after temozolomide remedy Luciana Andrade1; Andreia H. Otake2; Silvia Cardim1; Mariana Ikoma1; Felipe Silva1; Roger Chammas1Instituto do Cancer do Estado de Sao Paulo-ICESP, Sao Paulo, Brazil; ICESP FMUSP, Sao Paulo, Brazil; 3ICESP FMUSP, Sao Paulo, BrazilPS08.Exosomes boost SH-SY5Y neuroblastoma cells radioresistance by activating the AKT survival pathway Flavia Tortolici1; Anna Giovanetti2; Giulia Carrozzo3; Francesca Mastrostefano4; Stefano Rufini4 Department of Biology University of Rome “Tor Vergata”, Rome, Italy; Technical Unit for Radiation Biology and Human Overall health ENEA CR Casaccia, Roma, Italy; 3Department of Biology University of Rome “Tor Vergata, Rome, Italy; 4Department of Biology University of Rome “Tor Vergata”, Rome, Italy1Background: Extracellular vesicles (EVs) are emerging as a key players in intercellular communication. It has been shown that tumour cells EP Inhibitor MedChemExpress secrete huge amounts of EVS that may be taken up by malignant and stromal cells. A number of groups have demonstrated that EVs shed by tumour cells can induce resistance to therapy advertising tumour growth. Depending on that, our target will be to investigate if EVs secreted by melanoma cells in response to chemotherapy can modulate tumour development and progression. Approaches: Human melanoma cell lines have been treated with temozolomide (TMZ) and EVs secreted beneath these conditions have been purified from cell media following ultracentrifugation. EVs quantification was determined utilizing Nanosight NT LM10. The presence of Annexin V, CD9 and CD63 were determined making use of a flow cytometry. For macrophage polarization research, murine macrophages have been incubated with LPS and interferon gamma or IL4 in the presence of EVs derived from TMZ or vehicle melanoma treated cells to induce M1 and M2 polarization respectively. Right after 24 h, M1 and M2 gene expression have been determined by qPCR. For in vivo studies, human melanoma cells admixed with EVs derived from TMZ or vehicle treated cells had been injected s.c. in nude mice. Tumour development was measured with a caliper. Statistical analysis was performed employing GraphPad Prism. Outcomes: Our CB1 Antagonist Storage & Stability findings showed a significant raise in EVs secreted by human melanoma cell lines in response to TMZ remedy. Nanotracking evaluation revealed that the majority of EVs range from one hundred to 200 nm in size, comprising each exosome and microvesicles which have been good for CD9, CD63 and Annexin V. We observed that EVs shed by melanoma cells just after TMZ therapy modulate macrophage phenotype by skewing macrophage activation towards the MSaturday, 05 Mayphenotype as demonstrated by the.