The mean fluorescence intensity indicates superoxide levels. Student’s t-tests were performed to determine differences between samples. A value of p<0.05 was considered statistically significant. Results: A decreased intracellular superoxide production was observed in leucocytes from advanced HCC patients compared to patients without HCC (1739±105 versus 3145±443 fluorescence arbitrary units (FAU), p<0.05, respectively). The decreased production of intracellular superoxide was observed in monocytes, neutrophils and lymphocytes. Analysis of mitochondrial super-oxide formation with MitoSOX showed an increased (p<0.05) production of mitochondrial superoxide in leucocytes from HCC patients

that was observed in neutrophils (2667±407 vs 1266±84 FAU, p<0.05) and monocytes (3807±514 vs 2762±195 FAU, p<0.05) but not in lymphocytes (1827±231 vs 1977±160

PD0325901 purchase FAU, P>0.05). Conclusions: The results demonstrate a reverse effect of HCC on intracellular and mitochondrial superoxide levels. Leucocytes from patients with advanced HCC showed a decrease in intracellular superoxide and an increase in mitochondrial superoxide. These changes in super-oxide production in HCC patients could modulate immune responses through effects on signal transduction cascades. The determination of superoxide levels in leucocytes with FACS analysis might have value in the prediction of response to treatment in HCC patients. Disclosures: The following people have nothing to disclose: Paloma Lluch, Laia Navarro Lopez, Gloria Segarra, Guadalupe Herrera, Maria Mora, Joan Tosca, Miguel Angel Serra, Pascual Medina Background Stemness in cancer is currently of great interest GDC-0449 supplier as it can be used to predict prognosis of hepatocellular carcinoma (HCC). We recently proposed an HCC classification system defined by the stem cell markers epithelial cell adhesion molecule (EpCAM) and α-fetoprotein (AFP) to identify HCC subtypes closely related to certain liver lineages with distinct prognosis (Yamashita et al, Gastroenterology 2009). Here, we evaluated the utility of determining serum Dickkopf-1 (DKK-1) levels, encoded by DKK1, a gene activated by Wnt signaling Reverse transcriptase and co-regulated

with EPCAM, for the diagnosis of HCC with stem cell features. Material and Methods Patients diagnosed with HCC at the Liver Center, Kanazawa University Hospital, Japan from 2005 to 2012 were enrolled. We measured serum DKK-1 levels using the human DKK-1 ELISA kit (Uscn Life Science Inc.). Hepatic stem cell-like (HpSC-) and mature hepatocyte-like (MH-) HCCs were defined as previously described (Yamashita et al, Cancer Research 2008). Clinicopathological characteristics were determined and analyzed statistically in relation to serum DKK-1 concentrations using Kaplan-Meier survival analyses with log-rank tests, Cox proportional hazards models, Fisher’s exact tests, and logistic regression models. Results The study included 357 HCC patients.