On the other hand, the hepatic expression of the Sterol Regulatory Element Binding Protein-1c, that is the transcription factor activating all genes required for lipogenesis, was slightly reduced by CDAA treatment SLV 320 without major modifications induced by CCl4 administration in our experimental model. Furthermore, the Carbohydrate-Responsive Element-Binding Protein, a key element of glucosemediated stimulation of lipogenesis progressively decreased in CDAA-treated animals, independently from CCl4 administration, starting at 1 month. These data indicate that the development of steatosis in CDAA model was not associated with an increased lipogenesis. Thus, we analyzed the pathways related to FA oxidation, finding a decreased expression of both ACOX-1 and CPT1A observed in CDAA+CCl4-treated animals at 1 months and in CDAA mice thereafter, indicates that reduced fatty acid oxidation could be one of the mechanisms of hepatic steatosis. In this study, we have provided evidences that CDAA diet induces peripheral insulin resistance already in the first month after treatment, and this was associated to the pathological spectrum of NAFLD, including NASH and HCC. Peripheral insulin resistance is a primary feature of NAFLD/NASH, and is probably one of the main co-factors involved in the worsening of the disease. Thus the novelty of our results is the demonstration that the CDAA+CCl4 model determines peripheral insulin resistance, NAFLD and its progression to HCC. Using this novel experimental PNU 22394 hydrochloride approach we observed: a) development of peripheral insulin resistance already after 1 month; b) entire spectrum of lesions ranging from simple steatosis to NASH and HCC; c) development of HCC after 9 months of treatment in all mice; d) association of HCC development to increased extracellular matrix deposition; e) significant modification of oncogenic genes expression already after 3 months of treatment. Thus, this experimental model is able to guarantee in 9 months the development of HCC in almost 100% of animals and to early resemble the main features of the progression from NAFLD to NASH and HCC. In the majority of human cases, HCC arises in patients with advanced chronic liver injury and/or cirrhosis. NAFLD, which is present in up to 90% of all obese persons and in up to 70% of persons with type 2 diabetes, is a recognized risk factor for HCC, that may develop in NASH in the absence of cirrhosis. However, the study of the molecular mechanisms linking steatosis development to chronic liver injury and HCC is hampered by the lack of adequate experimental models that often do not resemble the human situation, either are not associated to a significant development of chronic liver injury or lead to a cachectic phenotype that does not allow a long period of observation, as needed for carcinogenesis.