5.1 cell transfection) using siRNA and a PKR expression plasmid, and then assessed cancer-related genes by real-time PCR and Western blotting. Cell lines were further analyzed using wound healing

and MTS (proliferation) assays. The modulation of genes by PKR was also assessed in 34 specimens of human HCC. Results: The expression of c-Fos and c-Jun genes was altered in parallel by PKR. An increase in PKR resulted in the upregulation of phosphorylated c-Fos and c-Jun that was related to levels of phosphorylated Erk1/2 and JNK1, namely the MAPK pathway, which is associated with cell proliferation. We therefore assessed cell proliferation in mono-layer wound-healing experiments. We found that JFH1 and H77s cells recovered more slowly buy GDC-0068 after PKR knockdown than controls. Cell proliferation determined by MTS assays significantly decreased and increased after PKR knockdown and PKR upregulation, respectively, and cell proliferation

Wnt inhibitor was dependent on PKR-modulated c-Fos and c-Jun expression. We also confirmed the coordinate expression of c-Fos and c-Jun with PKR in human HCC specimens with HCV infection. The amounts of c-Fos and c-Jun and their phosphorylation levels were increased in specimens with high levels of PKR expression. Conclusions: The activities of c-Fos and c-Jun were upregulated by PKR through the activation of Erk1/2 and JNK1, respectively. Such modulations resulted in HCC cell proliferation with HCV infection. These findings suggest that PKR-related proliferation pathways could Ixazomib cell line serve as an attractive therapeutic target. Disclosures: Raymond T. Chung – Advisory Committees or Review Panels: Idenix; Consulting: Enanta; Grant/Research Support:

Gilead, Merck, Mass Biologic, Gilead The following people have nothing to disclose: Takao Watanabe, Yoshio Tokumoto, Masashi Hirooka, Masanori Abe, Morikazu Onji, Yoichi Hiasa Hepatocellular carcinoma (HCC) is the third most lethal cancer worldwide, but molecular mechanisms of its pathogenesis are not well understood. Recent studies suggest that extracellular ATP-mediated activation of P2Y2 purinergic receptor induces hepatocyte proliferation in response to partial hepatectomy and ATP treatment alone was sufficient to induce hepatocyte proliferation in vitro. The purpose of this study was to characterize extracellular nucleotide effects on HCC cell proliferation and to examine the role of P2 purinergic signaling in the pathogenesis of HCC in patients and Mst1/2-/-, a mouse model of HCC. Hypothesis: Dysregulation of purinergic signaling facilitates aberrant cell proliferation underlying hepatocellular carcino-genesis. Methods. HCC human-derived Huh7 cells, maintained in serum free media for 24h, were treated with ATPγS, or ADP (100μM) for different time intervals. SP600125 pretreatment was used to inhibit c-Jun N-terminal Kinase (JNK) signaling. Western blotting, qRT-PCR and 5-Bromo-2′-deoxy-uridine (BrdU) incorporation analysis were done.