Reactive Oxygen Species Induce Epigenetic Instability through the Formation of 8-Hydroxydeoxyguanosine in Human Hepatocarcinogenesis
ABSTRACT Chronic hepatitis C (CHC) triggers oxidative stress and contributes to the emergence of hepatocellular carcinoma (HCC). We previously reported that tumor suppressor gene (TSG) methylation is a critical factor during the early stages of hepatocarcinogenesis. In this study, we clarify the association between oxidative stress and epigenetic alterations during hepatocarcinogenesis. We examined DNA oxidation and methylation profiles in 128 liver biopsy samples from CHC patients. The DNA oxidation and methylated TSG numbers were quantified using immunohistochemical analysis of 8-hydroxydeoxyguanosine (8-OHdG) and quantitative PCR for 11 TSGs, respectively. The quantitative chromatin immunoprecipitation-PCR (ChIP-qPCR) assay in HepG2 and fetal liver Hc cells treated with H2O2 was used to quantify trimethyl-H3K4, acetylated-H4K16 (an active chromatin marker), trimethyl-H3K27 (a repressive chromatin marker) and 8-OHdG. We analyzed 30 promoters of 25 different TSGs by qPCR. The high levels of 8-OHdG was the only variable that was significantly associated with the increased number of methylated TSGs in CHC (p < 0.0001). The ChIP-qPCR revealed that after H2O2 treatment of the cell lines, the 8-OHdG-bound promoters showed a modification from an active chromatin (trimethyl-H3K4 and acetylated-H4K16 dominant) to a repressive chromatin (trimethyl-H3K27 dominant) status. We conclude that oxidative stress alters the chromatin status, which leads to abnormal methylation of TSGs, and contributes to hepatocarcinogenesis in CHC patients. © 2013 S. Karger AG, Basel.
- Digestive Diseases 11/2013; 31(5-6):405-7. DOI:10.1159/000355379 · 2.18 Impact Factor
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ABSTRACT: HBx is a multifunctional regulator that interacts with host factors to contribute to the development of hepatocellular carcinoma. In this study, to explore the co-localization of HBx and COXIII in HepG2 cells and to investigate the molecular mechanism of HBx in HepG2 cell growth promotion, we first constructed a HepG2 cell line stably expressing the HBx gene in vitro by lentivirus vectors. In addition, we found that HBx co-localized with the inner mitochondrial protein, COXIII, in HepG2 cells by confocal laser scanning microscopy. It led to changes of mitochondrial biogenesis and morphology, including upregulation of COXIII protein expression, increased cytochrome c oxidase activity and higher mitochondrial membrane potential. The upregulation of COX-2 caused by HBx through generation of mitochondrial reactive oxygen species promoted cell growth. Thus, we conclude that co-localization of HBx and COXIII leads to upregulation of COX-2 that promotes HepG2 cell growth. Such a mechanism provides deeper insights into the molecular mechanism of HBV-associated hepatocellular carcinoma.International Journal of Oncology 06/2014; 45(3). DOI:10.3892/ijo.2014.2499 · 3.03 Impact Factor
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ABSTRACT: Hepatocellular carcinoma (HCC) is a common cancer worldwide and develops against a background of chronic liver damage. A variety of HCC-related genes are known to be altered by genetic and epigenetic mechanisms. Therefore, information regarding alteration of the genetic and epigenetic profiles in HCC is essential for understanding the biology of this type of tumor. Methylation at CpG sites in gene promoters is known to affect the transcription of the corresponding genes. Abnormal regional hypermethylation is observed in the 5' region of several tumor suppressor genes (TSGs) in HCC, and this hypermethylation may promote carcinogenesis through the transcriptional inactivation of downstream TSGs. The DNA damage induced by oxidation is a trigger of abnormal DNA methylation and inactivation of TSGs through recruitment of the polycomb repressive complex to the promoter sequence. Thus, oxidative stress may be responsible for the emergence of HCC from chronic hepatitis and liver cirrhosis through the epigenetic alteration of TSGs. There have been several attempts to apply epigenetic information to the diagnosis and treatment of HCC. The predictive value of selected methylation events on survival in HCC patients has been reported, and the methylation profile of background liver could be associated with recurrence-free survival of HCC patients who have undergone hepatectomy. Another study detected methylated DNA from HCC cells in serum, and the circulating tumor DNA was regarded as a potential tumor marker. In addition, several trials of HCC therapy have targeted the epigenetic machinery and were based upon comprehensive analyses of DNA methylation of this type of tumor. Here, we present an overview of research regarding DNA methylation status in human HCC and describe the clinical application of epigenetic information to HCC.10/2014; 3(3-4):417-27. DOI:10.1159/000343860