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Oxidative stress indicated by elevated expression of Nrf2 and 8-OHdG promotes hepatocellular carcinoma progression

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Abstract

Reactive oxygen species (ROS) is excessively generated in tumors creating an oxidative stress in tumor microenvironment. We investigated hepatic expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and 8-hydroxydeoxyguanosine (8-OHdG) in hepatocellular carcinoma (HCC) patients, and asked if ROS epigenetically upregulated Nrf2 and enhanced aggressiveness in HCC cells. Expression of Nrf2 (n = 100) and 8-OHdG (n = 53) was remarkably increased in HCC tissues compared with the noncancerous hepatic tissues. Elevated expression of 8-OHdG was associated with poor survival in HCC patients. H2O2, as ROS representative, provoked oxidative stress in HepG2 cells, indicated by increased protein carbonyl content and decreased total antioxidant capacity. Nrf2 expression and 8-OHdG formation were markedly increased in the H2O2-treated cells compared with the untreated control. Co-treatment with antioxidants, tocopheryl acetate (TA) and S-adenosylmethionine (SAM) effectively attenuated expression of Nrf2 and 8-OHdG in H2O2-treated cells. HepG2 cells treated with H2O2 had significantly higher migration and invasion capabilities than the untreated control cells, and this aggressiveness was significantly inhibited by TA and SAM. Bisulfite sequencing revealed that CpG dinucleotides in Nrf2 promoter were unmethylated in the H2O2-treated cells similar to the untreated control. In conclusion, robust histological evidence of increased antioxidative response and oxidative DNA damage in human HCC tissues was demonstrated. Elevated oxidative DNA lesion 8-OHdG was associated with shorter survival. Experimentally, ROS enhanced Nrf2 expression, 8-OHdG formation and tumor progression in HCC cells. These effects were inhibited by antioxidants. Therefore, oxidative stress-reducing regimens might be beneficial to diminish the ROS-induced HCC progression.

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Acknowledgement

This study was supported by the Ratchadapiseksompoch Endowment Fund, Chulalongkorn University (CU-58-001-HR to C.B.). C.M. awarded the 90th Anniversary of Chulalongkorn University Scholarship. We thank Miss Suchittra Phoyen and Miss Pimkanya Morekrong for laboratory assistance. Gratitude to Prof. Wilai Anomasiri for kindly help, advice and support.

Funding

This study was also funded (to P.T.) by the National Research University (NRU59-026-HR) and the Thailand Research Fund (RTA5980008).

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Authors and Affiliations

  1. Departments of Biochemistry, Faculty of Medicine, Chulalongkorn University, Rama IV Rd., Bangkok, 10300, Thailand

    Chakriwong Ma-on, Patcharawalai Whongsiri, Surasit Suwannasin, Pisit Tangkijvanich & Chanchai Boonla

  2. Departments of Pathology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand

    Anapat Sanpavat

  3. Center of Excellence in Immunology and Immune-mediated Diseases, Departments of Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand

    Nattiya Hirankarn

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  1. Chakriwong Ma-on
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Correspondence to Chanchai Boonla.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Ma-on, C., Sanpavat, A., Whongsiri, P. et al. Oxidative stress indicated by elevated expression of Nrf2 and 8-OHdG promotes hepatocellular carcinoma progression. Med Oncol 34, 57 (2017). https://doi.org/10.1007/s12032-017-0914-5

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  • DOI: https://doi.org/10.1007/s12032-017-0914-5

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