HCC development is associated to peripheral insulin resistance in a mouse model of NASH.
NAFLD is the most common liver disease worldwide but it is the potential evolution to NASH and eventually to hepatocellular carcinoma (HCC), even in the absence of cirrhosis, that makes NAFLD of such clinical importance.


OBJECTIVE
we aimed to create a mouse model reproducing the pathological spectrum of NAFLD and to investigate the role of possible co-factors in promoting HCC.


METHODS
mice were treated with a choline-deficient L-amino-acid-defined-diet (CDAA) or its control (CSAA diet) and subjected to a low-dose i.p. injection of CCl4 or vehicle. Insulin resistance was measured by the euglycemic-hyperinsulinemic clamp method. Steatosis, fibrosis and HCC were evaluated by histological and molecular analysis.


RESULTS
CDAA-treated mice showed peripheral insulin resistance at 1 month. At 1-3 months, extensive steatosis and fibrosis were observed in CDAA and CDAA+CCl4 groups. At 6 months, equal increase in steatosis and fibrosis was observed between the two groups, together with the appearance of tumor. At 9 months of treatment, the 100% of CDAA+CCl4 treated mice revealed tumor versus 40% of CDAA mice. Insulin-like Growth Factor-2 (IGF-2) and Osteopontin (SPP-1) were increased in CDAA mice versus CSAA. Furthermore, Immunostaining for p-AKT, p-c-Myc and Glypican-3 revealed increased positivity in the tumors.


CONCLUSIONS
the CDAA model promotes the development of HCC from NAFLD-NASH in the presence of insulin resistance but in the absence of cirrhosis. Since this condition is increasingly recognized in humans, our study provides a model that may help understanding mechanisms of carcinogenesis in NAFLD.
Università Politecnica delle Marche, Department of Gastroenterology, Ancona, Italy.
Samuele
S
De Minicis
Università Politecnica delle Marche, Department of Gastroenterology, Ancona, Italy.
Laura
L
Agostinelli
Università Politecnica delle Marche, Department of Gastroenterology, Ancona, Italy.
Chiara
C
Rychlicki
Università Cattolica del Sacro Cuore, Policlinico Gemelli, Division of Endocrinology and Metabolic Disease, Rome, Italy; Diabetic Care Clinics, ACISMOM, Rome, Italy.
Gian Pio
GP
Sorice
Università Politecnica delle Marche, Department of Gastroenterology, Ancona, Italy.
Stefania
S
Saccomanno
Università Politecnica delle Marche, Department of Gastroenterology, Ancona, Italy.
Cinzia
C
Candelaresi
Università Cattolica del Sacro Cuore, Policlinico Gemelli, Division of Endocrinology and Metabolic Disease, Rome, Italy; Don Gnocchi Foundation Onlus, Milan, Italy.
Andrea
A
Giaccari
Università Politecnica delle Marche, Department of Gastroenterology, Ancona, Italy.
Luciano
L
Trozzi
Università Politecnica delle Marche, Department of Gastroenterology, Ancona, Italy.
Irene
I
Pierantonelli
Università Politecnica delle Marche, Department of Gastroenterology, Ancona, Italy.
Eleonora
E
Mingarelli
Università Politecnica delle Marche, Department of Gastroenterology, Ancona, Italy.
Marco
M
Marzioni
Università Cattolica del Sacro Cuore, Policlinico Gemelli, Division of Endocrinology and Metabolic Disease, Rome, Italy.
Giovanna
G
Muscogiuri
Institute of Clinical Physiology, Head of Cardiometabolic Risk Unit, Pisa, Italy.
Melania
M
Gaggini
Università Politecnica delle Marche, Department of Gastroenterology, Ancona, Italy.
Antonio
A
Benedetti
Institute of Clinical Physiology, Head of Cardiometabolic Risk Unit, Pisa, Italy.
Amalia
A
Gastaldelli
Università di Padova, Department of Medicine Anatomic Pathology Unit, Padova, Italy.
Maria
M
Guido
Università Politecnica delle Marche, Department of Gastroenterology, Ancona, Italy; Università Politecnica delle Marche, Obesity Center, Ancona, Italy.
Gianluca
G
Svegliati-Baroni
D000367:Age Factors; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000209:etiology*; D002796:Choline Deficiency; D004195:Disease Models, Animal; D005526:Food, Formulated; D007333:Insulin Resistance / Q000502:physiology*; D007335:Insulin-Like Growth Factor II / Q000378:metabolism; D008113:Liver Neoplasms / Q000209:etiology*; D051379:Mice; D065626:Non-alcoholic Fatty Liver Disease / Q000150:complications* / Q000503:physiopathology*; D053495:Osteopontin / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism
C489428:Myc protein, mouse; D016271:Proto-Oncogene Proteins c-myc; C506896:Spp1 protein, mouse; D053495:Osteopontin; D007335:Insulin-Like Growth Factor II