Gene expression profiling of two cell lines established from an oval-cell derived hepatic tumor: Toward the discovery of a functional signature underlying liver cancer progression

Abstracts / Digestive and Liver Disease 41 (2009) A1–A45

GENE EXPRESSION PROFILING OF TWO CELL LINES ESTABLISHED FROM AN OVAL-CELL DERIVED HEPATIC TUMOR: TOWARD THE DISCOVERY OF A FUNCTIONAL SIGNATURE UNDERLYING LIVER CANCER PROGRESSION A.C. Piscaglia a,∗,1 , N. Saulnier a,1 , V. Tesori b , M. Barba a , M. Campanale a , F. Purchiaroni a , G. Ianiro a , T. Galeotti b , G. Pani b , A. Gasbarrini a a

Inst. of Internal Medicine, Catholic University of Rome, Gemelli Hospital, Rome, Italy b Inst. of General Pathology, Catholic University of Rome, Gemelli Hospital, Rome, Italy Background and aims. We established several cancer cell lines (named LCSCs) from a new model of hepatocholangiocarcinoma, based upon induction of oval cell proliferation prior to carcinogen exposure. Aim of this study was to compare the gene expression profile of two of these lines: LCSC-5 and LCSC-2. Methods. LCSCs were observed daily and their immunophenotype was assessed on cytospins and chamber-slides. The clonogenic potential of LCSCs was evaluated by soft-agar colony formation assay, and their tumorigenicity was investigated upon transplantation into nude mice. The growth kinetics was assessed by MTT assay. Gene expression profiling was performed on Affymetrix platform. The cellular response to HGF was measured by Western blot analysis for c-Met, p-ERK, pS6, p-Akt, and by functional assays. Finally, the effects of a novel and promising anticancer drug, Sorafenib, were investigated on both cell lines. Results. Although the immunophenotype and morphology of the two lines were similar, LCSC-2 had higher proliferative capacity and clonogenicity in vitro; moreover, upon transplantation, LCSC-2 induced macroscopic tumors in less than 30 days, while LCSC-5 required approximately 80 days. Transcriptional analysis revealed that genes overrepresented in LCSC-2 were mainly associated with tumor progression/growth (S100A4, ALDH3A1, ARMCX2), drug resistance (ABCC1) and invasion/cytoskeleton-remodelling (TPM2, PDLIM1, CAPG). Conversely, several liver-associated transcripts (IL6ST, ZFP36, IGF2, CYB5, LISCH7) and genes commonly downregulated in invasive cancers (GRO1, CEACAM1, NFKBIA, MGP) were reduced in LCSC-2. Interestingly, the expression level of the HGF receptor c-Met and its downstream signaling cascade appeared downregulated in LCSC-2 cells, compared to LCSC-5. Sorafenib had cytotoxic effects and, at cytostatic concentrations, potentiated HGF responses in both cell lines, this effect being again less pronounced in LCSC-2, which was also less sensitive to the drug. Conclusion. LCSC-2 displayed a more aggressive, less differentiated, and more drug-resistant phenotype when compared to LCSC-5. The HGF/c-Met pathway may play a

causative role in this difference, likely by promoting hepatic differentiation. The gene expression analysis allowed the identification of a panel of genes underlying liver cancer invasiveness and aggressiveness and which might offer novel insights to study the effects of molecularly targeted agents against hepatic tumors. doi:10.1016/j.dld.2008.12.068 HCV INFECTED PATIENTS ARE “SPARED” FROM THE METABOLIC SYNDROME BUT NOT FROM INSULIN RESISTANCE S. Ballestri a,∗ , E. Violi a , A. Lonardo a , L.E. Adinolfi b , L. Carulli a , S. Lombardini a , F. Scaglioni a , M. Ricchi a , G. Ruggiero b , P. Loria a a

Università degli Studi di Modena e Reggio Emilia, Medicina Metabolica NOCSAE Modena, Italy b Università degli Studi di Napoli, Medicina Interna ed Epatologia, Naples, Italy Background. Nonalcoholic fatty liver disease (NAFLD) and chronic hepatitis C feature steatosis and insulin resistance (IR), conditions associated with metabolic syndrome (MS). However, while NAFLD is associated with the MS, little is known about the relationship between HCV and MS. Aims. To assess prevalence of MS and determinants of IR in patients with NAFLD compared with cases with chronic hepatitis C. Methods. We enrolled 93 consecutive patients with NAFLD, 97 with chronic hepatitis C genotypes 1,2 and 182 “healthy” controls without steatosis. The prevalence of MS was assessed by modified ATP III criteria and IR by the homeostasis model assessment (HOMA-IR). IR was defined as the 75th percentile of HOMA-IR of control subjects. Results. While the prevalence of IR was similar in NAFLD and HCV (70.0% and 78.7%), the prevalence of the MS was significantly higher in NAFLD than in HCV (28.1% vs. 4.4%) and in controls (5.6%). At multivariate analysis, IR was predicted by body mass index (OR 1.263, C.I. 1.078–1.480) and triglyceridaemia (OR 1.011, CI 1.002–1.020) in NAFLD and by gender (OR for female sex 0.297, C.I. 0.094–0.940) and fibrosis stage (O.R. 2.751, C.I. 1.417–5.340) in chronic hepatitis C. Conclusions. IR, independently associated with BMI and triglyceridaemia in NAFLD, gender and fibrosis in HCV chronic hepatitis, has a higher prevalence in NAFLD and chronic hepatitis C than in controls. However, the frequency of the MS in HCV infected patients, similar to that of controls, is significantly lower than that seen in NAFLD. The current diagnostic criteria of MS are more likely to “capture” patients with NAFLD than with chronic hepatitis C, although both groups are insulin resistant. doi:10.1016/j.dld.2008.12.069

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These authors contributed equally.

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