897 INCREASED INTERLEUKIN-17-PRODUCING CD4 T CELLS CORRELATE WITH DISEASE PROGRESSION IN HEPATOCELLULAR CARCINOMA PATIENTS

POSTERS density-dependent: in well-spread cells, hepaCAM is distributed to cell protrusions whereas, in confluent cells, hepaCAM is predominantly accumulated at the sites of cell-cell contacts on cell membrane. In addition, hepaCAM colocalizes with filamentous actin (F-actin) on the plasma membrane, which can be disrupted by depolymerization of F-actin. Sucrose density gradients demonstrate that hepaCAM partitions into the lipid rafts while dispersion of cholesterol in the lipid rafts alters the buoyancy of hepaCAM. Moreover, hepaCAM colocalizes with the lipid raft markers, caveolin-1 and fyn. We have recently found that the cytoplasmic domain of hepaCAM is cleaved through multi-signaling pathways. Functionally, hepaCAM not only increases cell spreading, delays cell detachment and enhances cell migration, but also reduces cell colony formation and inhibits cell growth. Interestingly, when the cytoplasmic domain is deleted, little change can be observed with regard to the cell surface localization and dimer formation. However, the mutants have significantly lost their involvement in cell-matrix adhesion, cell motility and cell growth control. Overall, these data indicate that hepaCAM is a new Ig-like cell adhesion molecule and a tumor suppressor. The cytoplasmic domain is essential to the functions of hepaCAM on both cell growth arrest and cell-matrix interactions. 897 INCREASED INTERLEUKIN-17-PRODUCING CD4 T CELLS CORRELATE WITH DISEASE PROGRESSION IN HEPATOCELLULAR CARCINOMA PATIENTS M. Shi, F. Shi, J.-Y. Zhang, L.-M. Chen, F.-S. Wang. Beijing Institute of Infectious Diseases, Beijing 302 Hospital, Beijing, China E-mail: [email protected] Backgroup and aims: Hepatocellular carcinoma (HCC), which is mostly associated with chronic HBV infection, is high postsurgical recurrence and extremely poor prognosis. Substantial evidence indicated that the inflammatory reaction at tumor site can promote tumor growth and progression. We have recently reported that T helper (Th) 17 cells (Th17) exhibit the potential to exacerbate liver damage during chronic HBV infection. It is still controversial that Th17 play a pivotal role in tumor immunity. In this study, we identified the feature of Th17 in HCC patients and evaluate their correlation with disease progression. Methods: Seventy-eight HCC patients, 20 liver cirrhosis patients (LC) and 20 healthy donors (HC) were enrolled in this study. The phenotypic and functional features and distribution of Th17 were analyzed by flow cytometric analysis and/or immunohistochemistric staining. Results: We found that the frequency of Th17 was markedly upregulated in HCC patients as compared with HC and LC individuals. Th17 frequency was significantly higher in tumor infiltrating lymphocyte (TIL) than in non-tumor-infiltrating lymphocyte (NIL). IL-17-positive cells were preferentially recruited in tumor tissue and mainly accumulated in the peritumoral area. The expression of CD95 and Ki67 on Th17 was up-regulated in peripheral Th17, however, down-regulated in the tumor Th17. The contents of IL-17, IL-4, IL-10, IL-23 and IFN-g were much higher in non-tumor than tumor. PD-1 was up-regulated in Th17 of HCC patients, and blocking PD-1 with anti-PD-1 antibodies could largely enhance IL-17-production. Furthermore, we found that the concentration of IL-17 positively correlated with levels of cytokines such as IL-23, IL-1b, IL-6, TNF-a, MCP-1 and IL-8. IL-17 could activate monocytes to produce proinflammatory cytokines including IL-1b, IL-6, IL-23 and TNF-a. Conclusion: Th17 was highly accumulated in TIL, especially in peritumor with impaired functions, which was correlated with HCC progression. This study extended our notion of Th17 role in tumor immunity.

898 CLOSE ASSOCIATION OF ARREST DEFECTIVE PROTEIN 1 MRNA OVEREXPRESSION WITH MICROVASCULAR INVASION IN HEPATOCELLULAR CARCINOMA J.H. Shim1 , Y.-H. Chung1 , J.A. Kim1 , D.-J. Yoo1 , Y.-J. Jin1 , D. Lee1 , D.D. Seo2 , S.H. Kim3 , S.H. Ryu4 , S.H. Yang3 , E. Yu5 , Y.J. Lee6 . 1 Internal Medicine, University of Ulsan College of Medicine, Asan Medical Center, 2 Internal Medicine, University of Inje College of Medicine, Sanggye Paik Hospital, 3 Internal Medicine, Korea Veterans’ Hospital, 4 Internal Medicine, University of Inje College of Medicine, Seoul Paik Hospital, 5 Pathology, 6 Hepato-Biliary and Pancreatic Surgery, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Republic of Korea E-mail: [email protected] Background and Aims: Arrest defective protein 1 (ARD1) stabilizes hypoxia-inducible factor-1a (HIF-1a) as an acetylase, which, in turn, plays significant roles in the proliferation of several cancer cells. In this study we evaluated the association of ARD1 mRNA overexpression with clinicopathologic characteristics as well as long-term clinical outcomes of patients with HCC. Methods: ARD1 mRNA levels were measured in HCC and surrounding liver tissues obtained from 94 patients treated with partial hepatectomy (HBV/HCV/NBNC, 75/4/15). They were quantified by RT-PCR using ABI PRISM 7700 Sequence Detection System (Applied Biosystems). All samples were tested in duplicate. A housekeeping gene, GAPDH mRNA is used for the normalization of ARD1 mRNA levels. We also classified the subjects into two groups based on the 2−DDCT Method (high expression [2−DDCT > 1] and low expression [2−DDCT < 1] groups), and then compared various clinical, radiological and histological characteristics of HCC patients between the two groups. Results: In all, the ARD1 mRNA expressions in HCC tissues were not significantly different from those in surrounding liver tissues [ARD1 mRNA/GAPDH mRNA (×103 ); 1.97±1.21 vs. 1.74±1.45; p = 0.2]. However, very interestingly, microvascular invasions were more commonly observed in HCC tissues of high ARD1 mRNA expression group (2−DDCT > 1; n = 38) than in those of low ARD1 mRNA expression group (2−DDCT < 1; n = 56). (32% vs. 14%; p < 0.05) There was no difference in age, gender, etiology, Child–Pugh class, the extent and type of tumor, a-fetoprotein level, EdmondsonSteiner grade, presence of liver cirrhosis or portal vein thrombosis, TNM stage, and CLIP score between the two groups. The 1-, 3- and 5-year recurrence-free survival rates of HCC patients in high ARD1 mRNA expression group were not different from those in low ARD1 mRNA expression group (59%, 53% and 34% vs. 71%, 56% and 46%, respectively; p = 0.5). Also, the overall survival rates were not different from each other between the two groups (76% vs. 73% at 5 years and 59% vs. 65% at 10 years; p = 1.0). Conclusions: Our data suggested that ARD1 might be involved in the process of microvascular invasion in patients with HCC. 899 MICRORNA EXPRESSION OF KERATIN 19 POSITIVE HEPATOCELLULAR CARCINOMAS INDICATES STRONG TUMOUR PROGRESSION AND INVASION O. Govaere1 , B. Spee1 , C. Verslype2 , R. Aerts3 , B. Topal3 , F. Nevens2 , T. Roskams1 . 1 Department of Morphology and Molecular Pathology, Katholieke Universiteit Leuven, 2 Department of Hepatology, 3 Department of Abdominal Surgery, University Hospital Gasthuisberg, Leuven, Belgium E-mail: [email protected] Background and Aims: Keratin (K) 19 positivity in hepatocellular carcinomas (HCCs) has been correlated with a higher recurrence and a shorter overall survival. This phenotype of HCCs also express metastasis markers and may derive from progenitor cells. Hitherto

Journal of Hepatology 2010 vol. 52 | S319–S457

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