Hepatic Iron Overload Induces Hepatocellular Carcinoma in Transgenic Mice Expressing the Hepatitis C Virus Polyprotein

Abstract

Background & Aims: Despite the evidence of hepatic iron overload in patients with chronic hepatitis C, it remains unknown if iron overload is related to hepatocarcinogenesis in this condition. The aim of this study was to determine whether iron overload contributes to development of hepatocellular carcinoma (HCC) in transgenic mice expressing the hepatitis C virus (HCV) polyprotein. Methods: Male C57BL/6 transgenic mice expressing the HCV polyprotein and nontransgenic littermates were fed an excess-iron diet or control diet. Mice in each group were assessed for altered liver morphology and function and the development of liver tumors. Results: Hepatic iron concentrations in mice fed the excess-iron diet were comparable to those of patients with chronic hepatitis C. There was no inflammation in transgenic and nontransgenic livers. Compared with mice in 3 other groups, transgenic mice fed the excess-iron diet showed marked hepatic steatosis including the centrilobular microvesicular type, ultrastructural alterations of the mitochondria and decreased degradation activity of fatty acid at 6 months, and greater hepatic content of lipid peroxidation products and 8-hydroxy-2′-deoxyguanosine at 12 months after initiation of feeding. The number of proliferating hepatocytes was significantly increased in mice fed the excess-iron diet but was not different between transgenic and nontransgenic mice. Hepatic tumors including HCC developed in 5 of 11 (45%) transgenic mice fed the excess-iron diet but not in mice in other groups at 12 months after initiation of feeding. Conclusions: Iron overload induces mitochondrial injury and increases the risk of HCC development in transgenic mice expressing the HCV polyprotein. Background & Aims: Despite the evidence of hepatic iron overload in patients with chronic hepatitis C, it remains unknown if iron overload is related to hepatocarcinogenesis in this condition. The aim of this study was to determine whether iron overload contributes to development of hepatocellular carcinoma (HCC) in transgenic mice expressing the hepatitis C virus (HCV) polyprotein. Methods: Male C57BL/6 transgenic mice expressing the HCV polyprotein and nontransgenic littermates were fed an excess-iron diet or control diet. Mice in each group were assessed for altered liver morphology and function and the development of liver tumors. Results: Hepatic iron concentrations in mice fed the excess-iron diet were comparable to those of patients with chronic hepatitis C. There was no inflammation in transgenic and nontransgenic livers. Compared with mice in 3 other groups, transgenic mice fed the excess-iron diet showed marked hepatic steatosis including the centrilobular microvesicular type, ultrastructural alterations of the mitochondria and decreased degradation activity of fatty acid at 6 months, and greater hepatic content of lipid peroxidation products and 8-hydroxy-2′-deoxyguanosine at 12 months after initiation of feeding. The number of proliferating hepatocytes was significantly increased in mice fed the excess-iron diet but was not different between transgenic and nontransgenic mice. Hepatic tumors including HCC developed in 5 of 11 (45%) transgenic mice fed the excess-iron diet but not in mice in other groups at 12 months after initiation of feeding. Conclusions: Iron overload induces mitochondrial injury and increases the risk of HCC development in transgenic mice expressing the HCV polyprotein. Hepatocellular carcinoma (HCC) is a common malignancy and its incidence is increasing both in Japan and in the United States.1El-Serag H.B. Mason A.C. Rising incidence of hepatocellular carcinoma in the United States.N Engl J Med. 1999; 240: 745-750Crossref Scopus (2736) Google Scholar Persistent hepatitis C virus (HCV) infection is a major risk factor for the development of HCC. Approximately 80% of Japanese HCC patients are also diagnosed with HCV-associated cirrhosis or chronic hepatitis C.2Kiyosawa K. Tanaka E. Sodeyama T. Hepatitis C virus and hepatocellular carcinoma.in: Reesink H.W. Hepatitis C virus. Karger, Basel1998: 161-180Crossref Scopus (43) Google Scholar Nevertheless, the mechanisms underlying HCV-associated hepatocarcinogenesis are incompletely understood. Numerous studies have shown that oxidative stress is present in chronic hepatitis C to a greater degree than in other inflammatory liver diseases.3Barbaro G. Di Lorenzo G. Asti A. Ribersani M. Belloni G. Grisorio B. 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Mitochondrial injury, oxidative stress, and antioxidant gene expression are induced by hepatitis C virus core protein.Gastroenterology. 2002; 122: 366-375Abstract Full Text Full Text PDF PubMed Scopus (830) Google Scholar Besides the directly induced generation of ROS by the HCV proteins, chronic inflammation with production of proinflammatory cytokines has the potential to deliver an additional burden of ROS. These multiple sources of ROS production create a procarcinogenic environment under which chromosomal damage is likely to occur.See editorial on page 2229. See editorial on page 2229. Hepatic iron accumulation has been shown to be present in patients with chronic hepatitis C,6Kageyama F. Kobayashi Y. Kawasaki T. Toyokuni S. Uchida K. Nakamura H. Successful interferon therapy reverses enhanced hepatic iron accumulation and lipid peroxidation in chronic hepatitis C.Am J Gastroenterol. 2000; 95: 1041-1050Crossref PubMed Google Scholar, 7Farinati F. Cardin R. De Maria N. 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These mice showed no evidence of the in levels in nontransgenic and transgenic mice fed the excess-iron diet was to the degree of marked centrilobular microvesicular steatosis in transgenic mice fed the excess-iron of the mitochondria in mice of each group at 6 months after the of iron ultrastructural alterations of the mitochondria were in mice other than transgenic mice fed the excess-iron mitochondria were in transgenic mice fed the control diet. transgenic mice fed the excess-iron diet showed or the mitochondrial ultrastructural alterations of the mitochondria were shown in transgenic mice fed the excess-iron diet. transgenic mice fed the excess-iron diet steatosis centrilobular microvesicular steatosis and ultrastructural alterations of mitochondria mitochondrial in the transgenic mice fed in of acid acid is fatty the of the of and fatty are in the of fatty and PubMed Scopus (170) Google Scholar Compared with mice in the 3 other groups, transgenic mice fed iron showed significantly decreased in of acid These that iron and HCV mitochondrial Mitochondrial and ultrastructural alterations are likely to oxidative stress mitochondria are the of production of ROS. 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