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Involvement of copper in the liver tumorigenesis
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Abstracts / Digestive and Liver Disease 48S (2016) e18–e41
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T-31
INVOLVEMENT OF COPPER IN THE LIVER TUMORIGENESIS
SAFETY AND EFFICACY OF SOFOSBUVIR-BASED ANTIVIRAL THERAPY IMMEDIATELY AFTER LIVER TRANSPLANTATION: A MATCHED CASE–CONTROL ANALYSIS
C. Porcu 1 , L. Antonucci 1 , B. Barbaro 2 , M. Arciello 1 , C. Viscomi 1 , C. Balsano 2 1
Laboratory of Molecular Virology and Oncology, Francesco Balsano Foundation, Rome, Italy 2 Institute of Molecular Biology and Pathology (IBPM) – CNR, Rome, Italy Background: Hepatocellular carcinoma (HCC) represents the third most frequent cause of cancer death. The molecular mechanisms underlying the pathogenesis of HCC are still under investigation. Biometals and in particular copper (Cu), are emerging as important regulators of several physiological processes and alteration of their homeostasis is involved in several pathologies. Copper metabolism results significantly altered in neoplastic diseases: its concentration correlates with tumour incidence and burden, malignant progression, and recurrence in a variety of human cancers, including HCC. In particular, high serum and tissue levels of copper were found in liver cancer patients. Aim: We aimed to evaluate how the copper leads to liver tumorigenesis. Methods: We treated HepaRGTM cells, a human immortalized hepatic progenitor cell lines, with different copper sulfate (CuSO4 ) concentrations (5-100 M). The hepatoma cell line, HepG2 and HuH7.5, were treated with copper chelator Tetrathiomolybdate (TTM) (1-200 M). We also evaluated the viability by MTS assay, gene expressions by RT-PCR and protein levels by western blot. Results: An exposure to CuSO4 20 M for 6 days triggered an increase of HepaRG viability of about 50%. At the same time we appreciated an upregulation of genes involved in the promotion of the cell cycle (e.g. pcna, cyclinB, cyclinD1, cyclinE). Interestingly, the CuSO4 treatment promoted a marked increase of both mRNA and protein of Myc. The role of copper in modulating myc expression was strengthened by its down-regulation in HepG2 cells treated with a copper chelator TTM. Moreover, TTM treatment caused a reduction of cell viability and an up-regulation of the cell cycle inhibitor, p21. Conclusion: The copper seems to have a important role in inducing cancer phenotype, increasing cell proliferation and expression of different oncogenes, e.g. myc. Future investigations will be focused on the potential anti-cancer effects of natural polyphenols, e.g. Oleuropein, which we demonstrated to be able to chelate copper.
M. Sacco 1 , S. Martini 1 , D. Arese 1 , S. Strona 1 , F. Tandoi 2 , D. Cocchis 2 , S. Mirabella 2 , G. Rizza 2 , A. Ottobrelli 1 , M. Salizzoni 2 , M. Rizzetto 1 , G. Saracco 1 , R. Romagnoli 2 1 Gastrohepatology Unit, Department of Medical Sciences, A.O.U. Città della Salute e della Scienza di Torino, Turin, Italy 2 Liver Transplant Center, General Surgery Unit, Department of Surgical Sciences, A.O.U. Città della Salute e della Scienza di Torino, Turin, Italy
Introduction and aims: In Curry’s study, sofosbuvir + ribavirin prevented HCV recurrence after liver transplantation (LT) in 96% of patients with undetectable viremia for >30 days but only in 36% of patients who were negative for <30 days. We aimed to evaluate in patients still positive or negative for <30 days at LT safety and efficacy of a pre-emptive therapy immediately after LT. Methods: Between 07/2014 and 10/2015, in Turin LT Centre 37 HCV cirrhotic patients treated with sofosbuvir 400 mg daily and weight-based ribavirin underwent LT; 13 were still HCVRNA positive or negative for <30 days and continued antiviral therapy immediately after LT, for at least 12 weeks. Each patient was matched with 2 HCV recipients transplanted in our Centre before the advent of direct antivirals, between 12/2013 and 06/2014. Results: Treated group was similar to untreated one for recipient age (57.7 vs 54 years, p = .34), hepatocellular carcinoma prevalence (69% vs 54%, p = .50), MELD at LT (12 vs 13, p = .54), donor age (75 vs 63, p = .15), suboptimal graft quality (77% vs 54%, p = .30). Looking at the first 3 months after LT, the two groups did not differ for median hospitalization stay (15 vs 12 days, p = .35), ICU stay (4 vs 3 days, p = .29); acute rejection (0 vs 2 patients, p = .54); death (1 vs 1 patient, p = 1), however more treated patients showed ribavirininduced anemia and underwent blood transfusion (6 vs 3 patients, p = .04). At week 1, 2, 4, 8 and 12 post-LT, platelets count, INR, GGT, eGFR, tacrolimus daily dose and trough blood levels were similar in the two groups; transaminase remained instead significant higher in untreated patients. Until now, 10 patients achieved SVR12 and 2 are negative, still on treatment. Conclusions: Sofosbuvir + ribavirin therapy was safe and well tolerated immediately after LT, the only concern being ribavirinrelated anemia.
http://dx.doi.org/10.1016/j.dld.2015.12.080 http://dx.doi.org/10.1016/j.dld.2015.12.081
Abstracts / Digestive and Liver Disease 48S (2016) e18–e41
T-30
T-31
INVOLVEMENT OF COPPER IN THE LIVER TUMORIGENESIS
SAFETY AND EFFICACY OF SOFOSBUVIR-BASED ANTIVIRAL THERAPY IMMEDIATELY AFTER LIVER TRANSPLANTATION: A MATCHED CASE–CONTROL ANALYSIS
C. Porcu 1 , L. Antonucci 1 , B. Barbaro 2 , M. Arciello 1 , C. Viscomi 1 , C. Balsano 2 1
Laboratory of Molecular Virology and Oncology, Francesco Balsano Foundation, Rome, Italy 2 Institute of Molecular Biology and Pathology (IBPM) – CNR, Rome, Italy Background: Hepatocellular carcinoma (HCC) represents the third most frequent cause of cancer death. The molecular mechanisms underlying the pathogenesis of HCC are still under investigation. Biometals and in particular copper (Cu), are emerging as important regulators of several physiological processes and alteration of their homeostasis is involved in several pathologies. Copper metabolism results significantly altered in neoplastic diseases: its concentration correlates with tumour incidence and burden, malignant progression, and recurrence in a variety of human cancers, including HCC. In particular, high serum and tissue levels of copper were found in liver cancer patients. Aim: We aimed to evaluate how the copper leads to liver tumorigenesis. Methods: We treated HepaRGTM cells, a human immortalized hepatic progenitor cell lines, with different copper sulfate (CuSO4 ) concentrations (5-100 M). The hepatoma cell line, HepG2 and HuH7.5, were treated with copper chelator Tetrathiomolybdate (TTM) (1-200 M). We also evaluated the viability by MTS assay, gene expressions by RT-PCR and protein levels by western blot. Results: An exposure to CuSO4 20 M for 6 days triggered an increase of HepaRG viability of about 50%. At the same time we appreciated an upregulation of genes involved in the promotion of the cell cycle (e.g. pcna, cyclinB, cyclinD1, cyclinE). Interestingly, the CuSO4 treatment promoted a marked increase of both mRNA and protein of Myc. The role of copper in modulating myc expression was strengthened by its down-regulation in HepG2 cells treated with a copper chelator TTM. Moreover, TTM treatment caused a reduction of cell viability and an up-regulation of the cell cycle inhibitor, p21. Conclusion: The copper seems to have a important role in inducing cancer phenotype, increasing cell proliferation and expression of different oncogenes, e.g. myc. Future investigations will be focused on the potential anti-cancer effects of natural polyphenols, e.g. Oleuropein, which we demonstrated to be able to chelate copper.
M. Sacco 1 , S. Martini 1 , D. Arese 1 , S. Strona 1 , F. Tandoi 2 , D. Cocchis 2 , S. Mirabella 2 , G. Rizza 2 , A. Ottobrelli 1 , M. Salizzoni 2 , M. Rizzetto 1 , G. Saracco 1 , R. Romagnoli 2 1 Gastrohepatology Unit, Department of Medical Sciences, A.O.U. Città della Salute e della Scienza di Torino, Turin, Italy 2 Liver Transplant Center, General Surgery Unit, Department of Surgical Sciences, A.O.U. Città della Salute e della Scienza di Torino, Turin, Italy
Introduction and aims: In Curry’s study, sofosbuvir + ribavirin prevented HCV recurrence after liver transplantation (LT) in 96% of patients with undetectable viremia for >30 days but only in 36% of patients who were negative for <30 days. We aimed to evaluate in patients still positive or negative for <30 days at LT safety and efficacy of a pre-emptive therapy immediately after LT. Methods: Between 07/2014 and 10/2015, in Turin LT Centre 37 HCV cirrhotic patients treated with sofosbuvir 400 mg daily and weight-based ribavirin underwent LT; 13 were still HCVRNA positive or negative for <30 days and continued antiviral therapy immediately after LT, for at least 12 weeks. Each patient was matched with 2 HCV recipients transplanted in our Centre before the advent of direct antivirals, between 12/2013 and 06/2014. Results: Treated group was similar to untreated one for recipient age (57.7 vs 54 years, p = .34), hepatocellular carcinoma prevalence (69% vs 54%, p = .50), MELD at LT (12 vs 13, p = .54), donor age (75 vs 63, p = .15), suboptimal graft quality (77% vs 54%, p = .30). Looking at the first 3 months after LT, the two groups did not differ for median hospitalization stay (15 vs 12 days, p = .35), ICU stay (4 vs 3 days, p = .29); acute rejection (0 vs 2 patients, p = .54); death (1 vs 1 patient, p = 1), however more treated patients showed ribavirininduced anemia and underwent blood transfusion (6 vs 3 patients, p = .04). At week 1, 2, 4, 8 and 12 post-LT, platelets count, INR, GGT, eGFR, tacrolimus daily dose and trough blood levels were similar in the two groups; transaminase remained instead significant higher in untreated patients. Until now, 10 patients achieved SVR12 and 2 are negative, still on treatment. Conclusions: Sofosbuvir + ribavirin therapy was safe and well tolerated immediately after LT, the only concern being ribavirinrelated anemia.
http://dx.doi.org/10.1016/j.dld.2015.12.080 http://dx.doi.org/10.1016/j.dld.2015.12.081