Ligation-Mediated Polymerase Chain Reaction Detection of 8-Oxo-7,8-Dihydro-2 '-Deoxyguanosine and 5-Hydroxycytosine at the Codon 176 of the p53 Gene of Hepatitis C-Associated Hepatocellular Carcinoma Patients

A. Galli, A. Munnia, F. Cellai, M. Tarocchi, E. Ceni, F.J. van Schooten, R. Godschalk, R.W. Giese, M. Peluso*

*Corresponding author for this work

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Abstract

Molecular mechanisms underlying Hepatitis C virus (HCV)-associated hepatocellular carcinoma (HCC) pathogenesis are still unclear. Therefore, we analyzed the levels of 8-oxo-7,8-dihydro-2 '-deoxyguanosine (8-oxodG) and other oxidative lesions at codon 176 of the p53 gene, as well as the generation of 3-(2-deoxy-beta-d-erythro-pentafuranosyl)pyrimido[1,2-alpha]purin-10(3H)-one deoxyguanosine (M(1)dG), in a cohort of HCV-related HCC patients from Italy. Detection of 8-oxodG and 5-hydroxycytosine (5-OHC) was performed by ligation mediated-polymerase chain reaction assay, whereas the levels of M(1)dG were measured by chromatography and mass-spectrometry. Results indicated a significant 130% excess of 8-oxodG at -TGC- position of p53 codon 176 in HCV-HCC cases as compared to controls, after correction for age and gender, whereas a not significant increment of 5-OHC at -TGC- position was found. Then, regression models showed an 87% significant excess of M(1)dG in HCV-HCC cases relative to controls. Our study provides evidence that increased adduct binding does not occur randomly on the sequence of the p53 gene but at specific sequence context in HCV-HCC patients. By-products of lipid peroxidation could also yield a role in HCV-HCC development. Results emphasize the importance of active oxygen species in inducing nucleotide lesions at a p53 mutational hotspot in HCV-HCC patients living in geographical areas without dietary exposure to aflatoxin B-1.
Original languageEnglish
Article number6753
Number of pages12
JournalInternational Journal of Molecular Sciences
Volume21
Issue number18
DOIs
Publication statusPublished - 1 Sep 2020

Keywords

  • 8-oxodg
  • adducts
  • cancer
  • hcc
  • hcv
  • inflammation
  • m(1)dg
  • malondialdehyde
  • methylation
  • oxidative dna-damage
  • p53
  • pathogenesis
  • ros
  • somatic mutations
  • tp53
  • METHYLATION
  • CANCER
  • SOMATIC MUTATIONS
  • 8-oxodG
  • PATHOGENESIS
  • M(1)dG
  • MALONDIALDEHYDE
  • OXIDATIVE DNA-DAMAGE
  • HCC
  • ADDUCTS
  • INFLAMMATION
  • ROS
  • HCV
  • TP53

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