Macroscopic and microscopic observations were made. Gene expression and intracellular signaling pathways were analyzed. Oxidative stress, antioxidant, and macrophage deletion treatments were also performed in parallel with Cygb siRNA or CYGB overexpressing vectors transfection. Results: Model 1: 25 or 0.05 ppm DEN treatment for 25 or 36 weeks induced liver tumor formation in 1 00 or 43% of Cygb-KO mice, respectively, Belnacasan research buy compared to 44 or

0% of WT mice. In KO mice, there was liver fibrosis, increased inflammatory gene expression and augmented oxidative stress. Model 2: with as little as 8 weeks of CDAA treatment, Cygb-KO mice showed marked steatohep-atitis, which resulted in advanced fibrosis at 16 weeks, compared MK-8669 manufacturer with the WT. Surprisingly, at 32 weeks, 100% of Cygb-KO mice of both genders developed liver cancer, compared to 0% in WT mice. At all time points, there was severe inflammation associated with activated liver cancer pathways in the Cygb-KO mice. Cygb-KO HSC (HSC isolated from Cygb-KO mice) and Cygb siRNA-transfected WT-HSC were both primed, as indicated by markedly increased

expression of αSma, collagen 1α1, Tnfα, Tgfβ1, Il-1β, Il-6, Ccl-2, Ccl-3, and Ccl-4 mRNA and superoxide production compared to controls. Oxidative stress and antioxidant defense PCR arrays showed altered expression of 31 genes involved in the metabolism of reactive oxygen species in Cygb-KO mice. N-acetyl cysteine treatment for 2 weeks eliminated the oxidative stress in CDAA-fed Cygb-KO mice in vivo, and in cultures of isolated Cygb-KO HSC. Macrophage deletion after 8 weeks of CDAA feeding reduced the inflammatory reaction, oxidative stress, and fibrosis in Cygb-KO mice. Moreover, Hep-G2 cells overexpressing CYGB showed decreased proliferation under hypoxia compared with plasmid controls, selleck chemicals llc as well as downregulated expression of fibrosis- and angiogenesis-related genes. Conclusion: Cygb deficiency promotes liver

cancer development via HSC priming and augmented inflammation, local fibrosis, and oxidative stress. Disclosures: The following people have nothing to disclose: Thuy T. Le, Yoshinari Matsumoto, Hoang Hai, Katsutoshi Yoshizato, Norifumi Kawada Objective Overexpression of platelet-derived growth factor-C (PDGF-C) in the liver of mice (Pdgf-c Tg) induces hepatic fibrosis, followed by the development of hepatocellular carcinoma. We identified differentially expressed miRNAs in Pdgf-c Tg mice and evaluated their functional relevance in the progression of hepatic fibrosis and the development of HCC. Materials and Method We used TaqMan® Array Rodent MicroRNA A Cards v2.0 containing 384 miRNA assays. miRNAs were knocked down by locked nucleic acid (LNA)-modified antisense oligonu-cleotides (LNA-antimiR) in immortalized human stellate cells, Lx-2. Pdgf-c Tg mice at 32 months of age were injected with LNA-antimiR-214 via the tail vein six times (50 μg each) with 3-week intervals by using Invivofectamine® 2.0.