Methods: Mouse liver AS were studied by electron microscopy. Transcriptome analysis in AS cell lines vs. normal liver sinusoidal endothelial cells (LSEC) from control and Notch1 K〇 mice without AS including gene set enrichement analysis (GSEA) were performed. In one AS cell line the effects of BGJ398 supplier treatment with increasing sorafenib concentrations (1-20 μM) were analyzed. Time-lapse microscopy of sorafenib exposed AS cells grown on matrigel was analyzed. Cell proliferation was monitored using the real-time cell analyser system xCELLigence. Cell viability and intracellular signalling were assessed by flow cytometry and western

blotting. Results: In the transitional zone between tumor and normal tissue, ultrastructural features varied from normal to dysplastic endothelial cells Belnacasan with prominent nucle-oli and Weibel Palade bodies. Transcriptome analysis showed massive changes in gene expression identifying FGFRs, TGFβ, met proto-oncogene, PlGF, and VEGF-A as potential drivers in malignant transformation of hepatic AS. Moreover, GSEA revealed that six of the top 20 upregulated chemical and genetic perturbation gene sets were related to myc targets (FDR<0. 25). C-myc is a downstream transcription factor target of the Raf/MEK/ERK pathway, which can be blocked

by sorafenib, a multikinase inhibitor. Timelapse imaging showed that sorafenib treatment dramatically reduced migration of AS cells. Differences in filopodia dynamics were significant (p=0. 0201) after 6 h with a decrease in filopodia丨 extensions. Sorafenib inhibited cell proliferation in a time and dose-dependant manner, whereas the number of apoptotic cells was only slightly elevated with increasing concentrations. In addition, sorafenib suppressed ERK phosphorylation and expression of Cyclin D2 in the AS cell line. Conclusion: We identified Notch1 as LSEC tumor suppressor gene

and established 上海皓元 three hepatic AS cell lines as a useful in vitro tool. Our data demonstrate antitumor activity of sorafenib in AS cells with potent inhibition of migration, filopodia formation, and cell proliferation, which support further evaluation of sorafenib as a novel treatment strategy. Disclosures: The following people have nothing to disclose: Sonja Rothweiler, Michael T. Dill, Luigi Terraciano, Zuzanna Makowska, Markus H. Heim, David Semela Background: Hepatitis C virus (HCV) has been reported to regulate cellular microRNAs (miRNAs). The HCV core protein is considered to be a potential oncoprotein in HCV-related hepatocellular carcinoma (HCV-HCC), but HCV core-regulated microRNAs are largely unknown. Our preliminary experiments revealed significantly down-regulated microRNA-152 (miR152) expression in HCV core protein-overexpressing HepG2 cells in comparison with the control Ad-EGFP infected HepG2 cells.