- Email: [email protected]
P96 ALDOSE REDUCTASE CONTRIBUTES TO HEPATOCARCINOGENESIS IN TRANSALDOLASE DEFICIENCY
POSTERS for hepatocyte proliferation, was markedly downregulated. In a mouse model of HCC xenograft, ERK5 silencing or administration of XMD8–92 significantly decreased tumor volume. These effects were associated with reduced cell proliferation, as indicated by lower BrdU incorporation. Upon reduction of ERK5 levels of activity in vivo, levels of c-Rel and of c-Jun, a proto-oncogene essential for cell proliferation, were reduced. Expression of MEF2, a known target in the ERK5 pathway, was also inhibited. Conclusions: We found that ERK5 regulates cell proliferation in HCC in vivo and in vitro, affecting the expression of different oncogenic targets, including c-Rel, which we identify for the first time as a target of ERK5. P95 EXPRESSIONS OF MICRORNAS DIFFER BETWEEN CIRRHOTIC AND TUMOUR-SURROUNDING CIRRHOTIC LIVER AS WELL AS BETWEEN HEPATOCELLULAR CARCINOMA AND FOCAL NODULAR HYPERPLASIA K. Schlachter1 , B. Gyongy ¨ osi ¨ 1 , M. Gyugos1 , G. Lendvai1,2 , K. Baghy3 , 1,2 1 1 Z. Schaff , A. Kiss . 2nd Department of Pathology, 2 MTA-SE Tumor Progression Research Group, 3 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary E-mail: [email protected] Background and Aims: The altered expressions of microRNAs (miRNA) may contribute to liver diseases. In the present study, we determined the expressions of 12 miRNAs – suggested to be up- or downregulated in hepatocellular carcinoma (HCC) – in different tumorous and non-tumorous liver tissues. Methods: Isolated RNA samples of 75 patients were analysed including 29 HCC and 16 tumour-surrounding cirrhosis (TSCirrhosis), as well as 20 cirrhotic liver tissue (Cirrhosis), 10 focal nodular hyperplasia (FNH) and 15 normal liver tissue (Normal). The levels of miRNAs were determined using TaqMan MicroRNA Assays. Results: miR-17–5p and miR-210 expressions were reduced, whereas miR-34a and miR-224 levels were increased in the disease groups compared with Normal (p < 0.02). miR-122 was elevated in FNH (p < 0.3), although it is usually reduced in liver diseases. miR223 was decreased in HCC compared with Normal and Cirrhosis. Similarly, miR-214 was reduced in HCC as compared with TSCirrhosis (p < 0.03). The expressions of miR-221 and miR-222 were elevated in TS-Cirrhosis and HCC when compared with Cirrhosis (p < 0.02). The expressions of miR-18a, miR-21 and miR-222 were reduced in FNH as compared with HCC (p < 0.01). In HCC, miR-18a, miR-222 and miR-223 were found to be increased from low to high tumour grade (p < 0.04). Conclusions: miRNA expressional differences were revealed between FNH and HCC, Cirrhosis and TS-Cirrhosis. Increased miR221/222 expression characterized HCC compared with Cirrhosis, whereas miR-221/222 were reduced and miR-224 were elevated in Cirrhosis as compared with Normal. Acknowledgement: This work was supported by grants OTKA 101435 and K108548 from the National Scientific Research Fund.
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P96 ALDOSE REDUCTASE CONTRIBUTES TO HEPATOCARCINOGENESIS IN TRANSALDOLASE DEFICIENCY Z. Oaks1,2 , R. Hanczko1 , M. Beckford1 , S.K. Chung3 , S.K. Landas4 , J.M. Asara5 , A. Perl1,2,6 . 1 Medicine, 2 Biochemistry & Molecular Biology, SUNY – Upstate Medical University, Syracuse, NY, United States; 3 Department of Anatomy, The University of Hong Kong, Hong Kong, Hong Kong; 4 Department of Pathology, SUNY – Upstate Medical University, Syracuse, NY, 5 Division of Signal Transduction, Beth Israel Deaconess Medical Center, Boston, MA, 6 Microbiology and Immunology, SUNY – Upstate Medical University, Syracuse, NY, United States E-mail: [email protected] Background and Aims: Transaldolase (TAL) is the rate limiting enzyme of the pentose phosphate pathway. TAL deficiency is a model for NAFLD, NASH, and hepatocellular carcinoma (HCC) and results in the accumulation of sugar phosphates, depletion of NADPH and glutathione, which underlie oxidative stress (J. Clin. Invest. 2009;119:1546–57). TAL-deficient mice exhibit the overexpression of the NADPH dependent enzyme aldose reductase (AR). Here, we investigated if the deletion of AR influences the depletion of NADPH and its consequences in TAL deficiency. Methods: Metabolites were extracted from TAL, AR, and TAL/AR double-knockout livers that were sacrificed with C57BL/6 controls, using 5 age-matched mice of each strain. 258 metabolites were detected by LC-MS/MS with selective reaction monitoring. Relative metabolite quantities were then analyzed using Metaboanalyst to identify metabolites and pathways that were significantly changed. 21 TAL/AR double-knockout mice were monitored up to 78 weeks of age for the development of NAFLD, NASH, and HCC. Results: 0/21 TAL/AR double-knockouts, 45/97 TAL knockouts, and 3/106 controls developed HCC. Double-knockouts were not significantly different from controls (p=1.0), but had significantly reduced HCC compared to TAL knockouts (p < 0.0001). AR knockout livers had minimal changes relative to control animals. TAL knockout livers showed accumulation of sedoheptulose 7-phosphate (S7P) and erythrose 4-phosphate (E4P) and depletion of NADPH. TAL/AR knockout livers exhibited increased S7P and E4P but NADPH levels were normalized. Conclusions: These data show that AR activation contributes to NADPH depletion and oxidative stress and its blockade prevents hepatocarcinogenesis in TAL deficiency. P97 NEW INSIGHTS IN HOXA13 AND HOTTIP ROLE AS PROGNOSTIC FACTORS IN HEPATOCELLULAR CARCINOMA L. Quagliata1 , M. Matter1 , S. Piscuglio1 , L. Tornillo1 , M. Heim2 , C. Cillo3 , L. Terracciano1 . 1 Molecular Pathology Unit, Institute of Pathology, University Hospital Basel, 2 Department of Biomedicine, Hepatology Laboratory, University of Basel, Basel, Switzerland; 3 Federico II University Medical School, Naples, Italy E-mail: [email protected] Background and Aims: Previously, we observed that among the transcriptional factors family of HOX-genes, HOXA13 is highly deregulated in hepatocellular carcinoma (HCC). HOTTIP, a recently described long non-coding RNA, is located at the 5’HOXA locus (in contiguity with HOXA13) and binds WDR5/MLL-complexes driving transcription along the entire HOXA locus. Lately, we reported HOTTIP/HOXA13 deregulation as a key feature in HCC development, controlling liver-cancer cells apoptosis. Finally, we outlined HOTTIP/HOXA13 as predictive markers of HCCpatients’ outcome and disease progression. Here, driven by ChromatinImmunoprecipitation (ChiP) data, we corroborate our findings by screening a large cohort of HCCs evaluating HOXA13 levels and selected metastasis-associated genes directly regulated by HOXA13.
Journal of Hepatology 2014 vol. 60 | S67–S214
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P96 ALDOSE REDUCTASE CONTRIBUTES TO HEPATOCARCINOGENESIS IN TRANSALDOLASE DEFICIENCY Z. Oaks1,2 , R. Hanczko1 , M. Beckford1 , S.K. Chung3 , S.K. Landas4 , J.M. Asara5 , A. Perl1,2,6 . 1 Medicine, 2 Biochemistry & Molecular Biology, SUNY – Upstate Medical University, Syracuse, NY, United States; 3 Department of Anatomy, The University of Hong Kong, Hong Kong, Hong Kong; 4 Department of Pathology, SUNY – Upstate Medical University, Syracuse, NY, 5 Division of Signal Transduction, Beth Israel Deaconess Medical Center, Boston, MA, 6 Microbiology and Immunology, SUNY – Upstate Medical University, Syracuse, NY, United States E-mail: [email protected] Background and Aims: Transaldolase (TAL) is the rate limiting enzyme of the pentose phosphate pathway. TAL deficiency is a model for NAFLD, NASH, and hepatocellular carcinoma (HCC) and results in the accumulation of sugar phosphates, depletion of NADPH and glutathione, which underlie oxidative stress (J. Clin. Invest. 2009;119:1546–57). TAL-deficient mice exhibit the overexpression of the NADPH dependent enzyme aldose reductase (AR). Here, we investigated if the deletion of AR influences the depletion of NADPH and its consequences in TAL deficiency. Methods: Metabolites were extracted from TAL, AR, and TAL/AR double-knockout livers that were sacrificed with C57BL/6 controls, using 5 age-matched mice of each strain. 258 metabolites were detected by LC-MS/MS with selective reaction monitoring. Relative metabolite quantities were then analyzed using Metaboanalyst to identify metabolites and pathways that were significantly changed. 21 TAL/AR double-knockout mice were monitored up to 78 weeks of age for the development of NAFLD, NASH, and HCC. Results: 0/21 TAL/AR double-knockouts, 45/97 TAL knockouts, and 3/106 controls developed HCC. Double-knockouts were not significantly different from controls (p=1.0), but had significantly reduced HCC compared to TAL knockouts (p < 0.0001). AR knockout livers had minimal changes relative to control animals. TAL knockout livers showed accumulation of sedoheptulose 7-phosphate (S7P) and erythrose 4-phosphate (E4P) and depletion of NADPH. TAL/AR knockout livers exhibited increased S7P and E4P but NADPH levels were normalized. Conclusions: These data show that AR activation contributes to NADPH depletion and oxidative stress and its blockade prevents hepatocarcinogenesis in TAL deficiency. P97 NEW INSIGHTS IN HOXA13 AND HOTTIP ROLE AS PROGNOSTIC FACTORS IN HEPATOCELLULAR CARCINOMA L. Quagliata1 , M. Matter1 , S. Piscuglio1 , L. Tornillo1 , M. Heim2 , C. Cillo3 , L. Terracciano1 . 1 Molecular Pathology Unit, Institute of Pathology, University Hospital Basel, 2 Department of Biomedicine, Hepatology Laboratory, University of Basel, Basel, Switzerland; 3 Federico II University Medical School, Naples, Italy E-mail: [email protected] Background and Aims: Previously, we observed that among the transcriptional factors family of HOX-genes, HOXA13 is highly deregulated in hepatocellular carcinoma (HCC). HOTTIP, a recently described long non-coding RNA, is located at the 5’HOXA locus (in contiguity with HOXA13) and binds WDR5/MLL-complexes driving transcription along the entire HOXA locus. Lately, we reported HOTTIP/HOXA13 deregulation as a key feature in HCC development, controlling liver-cancer cells apoptosis. Finally, we outlined HOTTIP/HOXA13 as predictive markers of HCCpatients’ outcome and disease progression. Here, driven by ChromatinImmunoprecipitation (ChiP) data, we corroborate our findings by screening a large cohort of HCCs evaluating HOXA13 levels and selected metastasis-associated genes directly regulated by HOXA13.
Journal of Hepatology 2014 vol. 60 | S67–S214