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882 TUMOR LYMPHOCYTIC INFILTRATE: A PREDICTOR OF DISEASE FREE SURVIVAL AFTER RESECTION FOR HCC
POSTERS Erlotinib or Sorafenib alone. These data provides the rationale to test the combination in early clinical trials. 882 TUMOR LYMPHOCYTIC INFILTRATE: A PREDICTOR OF DISEASE FREE SURVIVAL AFTER RESECTION FOR HCC E. Cariani1 , M. Pilli2 , A. Zerbini2 , A. Olivani2 , C. Rota1 , G. Pelosi2 , P. Soliani3 , E.M. Silini4 , C. Ferrari2 , G. Missale2 . 1 Laboratorio di Tossicologia, Ospedale Civile S. Agostino-Estense, Modena, 2 Malattie Infettive ed Epatologia, Azienda Ospedaliero-Universitaria di Parma, 3 Clinica Chirurgica e dei Trapianti d’Organo, 4 Istituto di Anatomia Patologica, Universit` a degli Studi di Parma, Parma, Italy E-mail: [email protected] Background and Aims: Recent studies have analyzed the influence of host immunity on cancer prognosis. Tumor-infiltrating CD3+ T cells are strongly associated with favourable prognosis in several solid malignancies. The main aim of this study was to test the prognostic value and phenotypic characteristics of tumor infiltrating lymphocytes (TIL) within resected HCC nodules. Methods: HCCs from 30 patients with a mean follow-up of 4.5 years were evaluated for morphology by histological parameters (grade, vascular invasion, type of growth, nuclear cytoplasmic ratio, semiquantitative evaluation of lymphomonocytic infiltrate, Betacatenin expression, etc.), for expression of immunologically-related (CD45, CD8, FoxP3, Granzyme-B, IFN-gamma, PD-1, PDL-1) as well as fetal and staminal genes (AFP, IGF2, VIM, CK7, eCadherin, EpCam) by mRNA analysis and finally for phenotype and function of T and NK cells on isolated lymphomononuclear cells (cytokine production upon mitogen situmulation) by staining with a large panel of monoclonal antibodies. Patients were divided according to disease recurrence before and after 12 months and significantly different parameters (Mann Whitney and chi-square for morphological scores) were further evaluated for their prognostic relevance (disease free survival – DFS) by Kaplan Meier curves. Results: CD3+ and CD4+ T-cells by flow cytometry and lymphomonocytic infiltrate by histological scoring were significantly more represented in patients with no disease recurrence at 12 months while larger HCC size and vascular invasion were significantly associated with disease recurrence. However, when Kaplan Meier curves were applied, CD4+ T-cells (p = 0.01) and lymphomonocytic infiltrate (p = 0.03) appeared as the only significant predictors of DFS. Interestingly there was a positive correlation (Spearman rank correlation test) between lymphomonocytic infiltrate and expression of fetal markers (IGF2 and AFP). Moreover, even if frequency of CD4CD25FoxP3 Treg cells did not significantly impact on DFS, it was inversely correlated with CD4+ and CD8+ T-cells function (IFN-gamma, TNF-alpha and IL-2 expression upon mitogen stimulation). Conclusions: Even if several histological and immunological parameters were evaluated, intratumoral lymphocytic infiltrate and in particular CD4+ cells were the main predictors of disease free survival. Further qualitative analysis of infiltrating T, NK and NKT cells will provide insights into the mechanisms of tumor immunity in patients with hepatocellular carcinoma. 883 THE PIVOTAL ROLE OF HEPATIC-STELLATE-CELLS IN THE IN-VIVO AND IN-VITRO HEPATOCELLULAR CARCINOMA MODELS N. Muhanna, L. Abu Tair, S. Doron, J. Amer, R. Safadi. Liver Unit, Gastroenterology Institute, Division of Medicine, Hadassah University Medical Center, Jerusalem, Israel E-mail: [email protected] Background: Hepatic-stellate-cells (HSCs) activation is the key element in hepatic-fibrosis and cirrhosis progression. Cirrhosis is a main risk factor for hepatocellular-carcinoma (HCC) development. However, direct HSCs role in HCC-progression is not fully understood. S344
Aims: To evaluate the homeostatic balance of the interactions between HCC/HSCs in early and advanced models of fibrosis stages. Methods and Results: In-vitro; human-HSCs (LX2-line) co-cultured with HCC cell-line (Hep3B) decreased alpha-feto-protein secretions (aFP) to medium. Prominent reduction of aFP-secretion obtained when LX2/Hep3B were co-cultured in same well of the “doublechamber-wells” device. This reduction however, was inhibited when LX2/Hep3B were co-cultured separately in different transwells of the device, suggesting a direct cellular-contact is necessary in decreasing tumor progression. In addition, LX2 pre-activation by HCV-lymphocytes or leptin prior to co-culture with Hep3B, further suppressed aFP-levels and caused Hep3B-phagocytosis inside HSCs. Results were significant in HCV-derived lymphocytes of F4-fibrosis grade as compared to F0–1 (P = 0.02). Confocal-microscopy and flow-cytometry demonstrated phagocytosis of HCC-cells inside the HSCs confirming their direct anti-tumor effects. In-vivo; HCC model was induced by intra-hepatic injections of HepB3-cells in male Nude-nu mice. Mice with carbon-tetra-chloride induced liverfibrosis prior to HCC injections (a model for advanced fibrosis) had the higher tumor size and aFP serum-levels as compared to nonfibrotic livers (P = 0.008). However, a significant reduction in tumor size and mass was achieved when liver-fibrosis was administered post Hep3B-injections; a model of early HSCs-activation. Conclusions: Cirrhosis, as end-result of HSCs-activation, indirectly provides a pro-HCC condition. However, in the in-vivo/in-vitro models, early HSCs activation also expresses direct anti-tumor effects by phagocytosis of tumor-cells. HSCs have pivotal effects in modulation of tumor activity that depend on the stage/phase of their activation. 884 CXCL1 RS4074 POLYMORPHISM INFLUENCES THE SUSCEPTIBILITY TO HEPATOCELLULAR CARCINOMA 2 H.D. Nischalke1 , M. Coenen1 , C. Berger1 , T. Muller ¨ , T. Berg2 , 1 1 C. Luda1 , B. Kramer ¨ , D. Schulte1 , C. Korner ¨ , J. Nattermann1 , T. Sauerbruch1 , U. Spengler1 . 1 Medizinische Klinik und Poliklinik I, Universit¨ at Bonn, Bonn, 2 Medizinische Klinik mit Schwerpunkt Hepatologie und Gastroenterologie, Charit´e, Universit¨ atsmedizin Berlin, Berlin, Germany E-mail: [email protected] Background and Aims: CXCL1 also known as growth regulated oncogene-alpha (GROa) is a chemokine that contributes to many protumorigenic processes such as inflammation and angiogenesis. This gene has also been reported to be over-expressed in various forms of cancer, such as bladder, ovarian, gastric and hepatocellular carcinomas. In the present study, we investigated the influence of the CXCL1 rs4074 polymorphism on the occurrence of hepatocellular carcinoma. Methods: The study involved 285 patients with HCC (163 HCV associated, 49 HBV associated, 53 ethyl-toxic induced and 20 cryptogenic HCCs) and 244 healthy controls. The CXCL1 rs4074 polymorphism was determined using a LightSNiP assay on the LightCycler system. Results: Distribution of CXCL1 genotypes (G/G, G/A, A/A) matched the Hardy-Weinberg equilibrium in each group and was 95 (33.3%), 143 (50.2%), 47 (16.5%) in patients with HCC and 109 (44.7%), 102 (41.8%), 33 (13.5%) in healthy controls. Thus, the frequency of the CXCL1 rs4074 A allele was significantly higher in patients with HCC (41.6%) than in healthy controls (34.4%) (p = 0.017, Odds ratio = 1.356, 95%-C.I.:1.056–1.741). Conclusion: The finding of an increased frequency of the CXCL1 rs4074 A allele in patients with HCC indicates carriage of an A allele to be a potential risk factor predisposing to hepatocellular carcinoma.
Journal of Hepatology 2010 vol. 52 | S319–S457
Aims: To evaluate the homeostatic balance of the interactions between HCC/HSCs in early and advanced models of fibrosis stages. Methods and Results: In-vitro; human-HSCs (LX2-line) co-cultured with HCC cell-line (Hep3B) decreased alpha-feto-protein secretions (aFP) to medium. Prominent reduction of aFP-secretion obtained when LX2/Hep3B were co-cultured in same well of the “doublechamber-wells” device. This reduction however, was inhibited when LX2/Hep3B were co-cultured separately in different transwells of the device, suggesting a direct cellular-contact is necessary in decreasing tumor progression. In addition, LX2 pre-activation by HCV-lymphocytes or leptin prior to co-culture with Hep3B, further suppressed aFP-levels and caused Hep3B-phagocytosis inside HSCs. Results were significant in HCV-derived lymphocytes of F4-fibrosis grade as compared to F0–1 (P = 0.02). Confocal-microscopy and flow-cytometry demonstrated phagocytosis of HCC-cells inside the HSCs confirming their direct anti-tumor effects. In-vivo; HCC model was induced by intra-hepatic injections of HepB3-cells in male Nude-nu mice. Mice with carbon-tetra-chloride induced liverfibrosis prior to HCC injections (a model for advanced fibrosis) had the higher tumor size and aFP serum-levels as compared to nonfibrotic livers (P = 0.008). However, a significant reduction in tumor size and mass was achieved when liver-fibrosis was administered post Hep3B-injections; a model of early HSCs-activation. Conclusions: Cirrhosis, as end-result of HSCs-activation, indirectly provides a pro-HCC condition. However, in the in-vivo/in-vitro models, early HSCs activation also expresses direct anti-tumor effects by phagocytosis of tumor-cells. HSCs have pivotal effects in modulation of tumor activity that depend on the stage/phase of their activation. 884 CXCL1 RS4074 POLYMORPHISM INFLUENCES THE SUSCEPTIBILITY TO HEPATOCELLULAR CARCINOMA 2 H.D. Nischalke1 , M. Coenen1 , C. Berger1 , T. Muller ¨ , T. Berg2 , 1 1 C. Luda1 , B. Kramer ¨ , D. Schulte1 , C. Korner ¨ , J. Nattermann1 , T. Sauerbruch1 , U. Spengler1 . 1 Medizinische Klinik und Poliklinik I, Universit¨ at Bonn, Bonn, 2 Medizinische Klinik mit Schwerpunkt Hepatologie und Gastroenterologie, Charit´e, Universit¨ atsmedizin Berlin, Berlin, Germany E-mail: [email protected] Background and Aims: CXCL1 also known as growth regulated oncogene-alpha (GROa) is a chemokine that contributes to many protumorigenic processes such as inflammation and angiogenesis. This gene has also been reported to be over-expressed in various forms of cancer, such as bladder, ovarian, gastric and hepatocellular carcinomas. In the present study, we investigated the influence of the CXCL1 rs4074 polymorphism on the occurrence of hepatocellular carcinoma. Methods: The study involved 285 patients with HCC (163 HCV associated, 49 HBV associated, 53 ethyl-toxic induced and 20 cryptogenic HCCs) and 244 healthy controls. The CXCL1 rs4074 polymorphism was determined using a LightSNiP assay on the LightCycler system. Results: Distribution of CXCL1 genotypes (G/G, G/A, A/A) matched the Hardy-Weinberg equilibrium in each group and was 95 (33.3%), 143 (50.2%), 47 (16.5%) in patients with HCC and 109 (44.7%), 102 (41.8%), 33 (13.5%) in healthy controls. Thus, the frequency of the CXCL1 rs4074 A allele was significantly higher in patients with HCC (41.6%) than in healthy controls (34.4%) (p = 0.017, Odds ratio = 1.356, 95%-C.I.:1.056–1.741). Conclusion: The finding of an increased frequency of the CXCL1 rs4074 A allele in patients with HCC indicates carriage of an A allele to be a potential risk factor predisposing to hepatocellular carcinoma.
Journal of Hepatology 2010 vol. 52 | S319–S457