Methods: we engineered Huh7.5 and HepG2 human hepatoma cell lines to express Omomyc under the control of a tetracycline inducible promoter.

PLX4032 order Then, we determined by microscope examination, MTS assay and cytofluorimetric analysis, morphology, proliferation and viability of mock transduced cells, of cells transduced with Omomyc without induction of its expression, and of cells induced to express Omomyc. Real-time PCR and Western blot analysis were used for determining the expression of Myc, PCNA (Proliferating Cellular Nuclear Antigen), CCD1 (Cyclin D1), Hedgehog and its target gene Gli1. Results: Microscope examination, MTS assay and FACS analysis clearly revealed that Omomyc expression is able to reduce human hepatocel-lular carcinoma cell proliferation up to 70% in 8 days upon doxycycline induction. Analysis of mRNA expression of genes involved in cell proliferation and the correspondent protein levels confirmed the Omomyc effect in halting tumor cell growth. In particular, the expression of Hedgehog and its target gene Gli1 were decreased after Omomyc induction. Conclusion: The Myc dominant negative miniprotein EPZ-6438 cost Omomyc is able to prevent HCC proliferation in vitro by specifically counteracting the Hedgehog-mediated signaling. We are now testing the outcome of this strategy in preventing tumor progression

in vivo in SCID mice injected with HCC cells expressing Omomyc and luciferase, in order to follow cancer growth by bioluminescence imaging. Disclosures: The following people have nothing to disclose: Barbara Barbaro, Cristiana Porcu, Gabriele Toietta, Roberta Maggio, Mauro Savino, Sergio Nasi, Clara Balsano Background & Aim: Hepatocytes play a crucial role in the homeostatic control of energy metabolism. When hepatocytes are exposed to an intolerably high amount of fat under overfed state, their toxic metabolites excessively

accumulate in the cells, often leading to the lipotoxic liver injury called nonalcoholic fatty liver disease (NAFLD). check details Although epigenetic mechanisms have been discussed for the maintenance of metabolic homeo-stasis, it is unclear how epigenetic factors selectively control gene expression for the metabolic adaptation. Lysine-specific demethylase 1 and 2 (LSD1 and LSD2) comprise the flavin-dependent amine oxidase family of histone demethylases. We have previously reported that LSD1 represses energy expenditure genes in adipose cells under lipogenic conditions, giving a hint that another flavin-dependent epigenetic factor, LSD2 may link environmental information to metabolic programming. This study was aimed to identify a key role of LSD2 in the homeostatic control of energy metabolism in hepatocytes. Methods: Transcriptome analysis and bioinformatic approach using gene set enrichment analysis (GSEA) was carried out to identify the target genes of LSD2 in LSD2-depleted HepG2 human hepatic cells.