METHODS: Colon carcinoma CT26 cells, expressing HSF1, HSP70 and HSP90, were used to assess shRNA-HSF1-nanoliposomal uptake, toxic-ity/efficiency by employing immunofluorescence, Q-PCR and protein analysis. In vitro sub-lethal heat experiments were performed to mimic radiofrequency ablation and this by using different time points and temperatures. An orthotopic murine model of coloncarcinoma cancer – liver metastasis was used to analyze HSF1 expression during tumour formation. Radiofre-quency ablation (RFA) in small animals was optimized Sotrastaurin clinical trial to investigate the HSF1-induced-and
related signaling pathways in the treatment of liver metastasis. RESULTS: shRNA-HSF1-nanolipo-somes were taken up by 99 procent of cells without inducing cytotoxicity. Sub-lethal heat treatment of 45 and 50 degrees Celsius induced p-ERK, p-AKT and HSF1-related proteins at different timepoints investigated and this coincided with a nuclear to cytosolic shift of HSF1, HSP70/90, AKT and ERK. Apoptosis was significantly induced only after 10 days post-heat treatment. In vivo, see more tumours highly expressed HSF1, HSP70/90, AURBK and p-ERK and p-AKT. Radiofrequency-ablated tumours showed an increase in HSF1 and HSP70/90 protein expression after 6 and 10 days post-RFA, suggesting the involvement of HSF1 during the process
of tumour recurrence. CONCLUSION: This study demonstrates that HSF1 is highly expressed in CRC liver metastasis and suggest its possible involvement in tumour recurrence after employing radiofrequency ablation. Disclosures: Claudio Amabile – Employment: H.S. Hospital Service SpA Nevio Tosoratti – Employment:
HS HOSPITAL SERVICE SpA Simone Cassarino – Employment: H.S. Hospital Service SpA Mark Kester – Stock Shareholder: those Keystone Nano Inc Massimo Pinzani – Advisory Committees or Review Panels: Intercept Pharmaceutical, Silence Therapeutic, Abbot; Consulting: UCB; Speaking and Teaching: Gilead, BMS The following people have nothing to disclose: Francesca Zanieri, Vinicio Car-loni, Sara Omenetti, Sriram Saravanan Shanmuga Velandy, Krista Rombouts Background/Aims: Hepatitis E virus (HEV), a hepatotropic virus has shown the property of self-assembly through its N-terminally truncated ORF2 (Nt-ORF2). Recent studies have demonstrated virus-like particles (VLPs) as a drug-delivery vehicle. In this study, we developed HEV-LPs generation system based on Huh7 cells and confirmed liver-specific penetration of the HEV-LPs. Moreover, we established a HEV-LP disassembly/reassembly system to charge therapeutic agents into the VLPs. Methods: In order to deliver CMV promoter derived-Nt-ORF2 gene into Huh7 cells, we used Bac-to-Bac system.