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Mariantonietta Pisaturo,1 Alessandra Di Fraia,1 Laura Occhiello,1 Carmine Minichini,1 Mario Starace,1 Valentina Iodice,2 Nunzia Farella,2 Maria Stanzione,1 Nicola Coppola1 1Infectious Diseases, Department of Mental Health and Public Medicine, University of Campania “L. Vanvitelli”, Naples, Italy; 2IX Infectious Disease Unit, AORN dei Colli, Naples, ItalyCorrespondence: Nicola CoppolaDepartment of Mental Health and Public Medicine, Section of Infectious Diseases, University of Campania Luigi Vanvitelli, Via L. Armanni 5, Naples, 80131, ItalyTel +39 081 5666223Fax +39 081 5666013Email nicola.coppola@unicampania.itBackground: The present paper evaluates the genetic variability of HCV in patients with hepatocellular carcinoma (HCC).Methods: Amino acid substitutions (aas) in NS3, NS5A and core regions were analyzed in 17 patients with HCC (Cases) and 13 without HCC (Controls), all naïve to DAAs. For the Cases, a sample of neoplastic liver tissue, non-neoplastic liver tissue and a serum sample were collected; for the Controls, a sample of liver tissue was collected. Sanger sequencing of three regions was performed using homemade protocols.Results: Phylogenetic trees showed that there was no difference in the virus populations in the three compartments analyzed for the three HCV regions in patients with HCC. Low variability and no difference between the Cases and Controls were observed in the core and NS5A regions; however, in the NS3 region, a higher variability was observed in the Cases. No difference was observed in the core region between Cases and Controls. In NS3, aa substitutions at positions 103 and 122 were more frequently found in Cases than Controls (in both cases 50% vs 9.1%, p< 0.05); moreover, aas in positions 32, 44 (p=0.035 for both), 79 (p=0.008) and 121 (p=0.018) were observed in the Cases and absent in the Controls. Finally, considering the NS5A region, aa substitutions at positions 37 and 54 were more frequently identified in the Cases than the Controls, but without statistical significance.Conclusion: These data may suggest a higher aa variability in patients with HCC than in those without, especially in the NS3 region.Keywords: hepatocellular carcinoma, HCV, viral variability, chronic hepatitis C, chronic infection C, core region, NS5A region, NS3 region, aa substitutions, phylogenesis