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Blocking of JB6 Cell Transformation by Tanshinone IIA: Epigenetic Reactivation of Nrf2 Antioxidative Stress Pathway

  • Research Article
  • Theme: Natural Products Drug Discovery in Cancer Prevention
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Abstract

Increasing numbers of natural products have been found to possess anticancer effects. Nuclear factor erythroid-2-related factor-2 (Nrf2) is a master regulator of the antioxidative stress response, and our previous studies found that epigenetic modification of the Nrf2 gene appears to be a critical mechanism. Salvia miltiorrhiza, a Chinese herbal medicine widely used in Asian countries, has been shown to possess anticancer and antioxidant effects. Tanshinone IIA (TIIA), an active component in S. miltiorrhiza, has been reported to activate Nrf2 pathway. The objective of this study was to investigate the epigenetic regulation of Nrf2 by TIIA in mouse skin epidermal JB6 cells and the functional consequences for cell transformation. TIIA was found to induce antioxidant response element-luciferase and upregulate the mRNA and protein levels of Nrf2 and Nrf2 downstream target genes HO-1 and NQO-1. TIIA decreased the colony formation of JB6 cells by approximately 80%. TIIA decreased the protein levels of DNMT1, DNMT3a, DNMT3b, and HDAC3 and inhibited the enzymatic activity of HDACs. Bisulfite genomic sequencing indicated that TIIA demethylated the first five CpGs in the promoter region of the Nrf2 gene. Chromatin immunoprecipitation assays showed that TIIA treatment increased the recruitment of RNA polymerase II at Nrf2 transcription start site but had limited effects on enrichment of Ac-H3 in Nrf2 promoter. Taken together, our results show that TIIA activates the Nrf2 signaling pathway and induces epigenetic demethylation of the CpGs of Nrf2. The epigenetic reactivation of the Nrf2 signaling pathway by TIIA could potentially contribute to the attenuation of JB6 cellular transformation and anticancer effects.

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ACKNOWLEDGMENTS

This study was supported by Grant 81102856 from the National Science Foundation of China to Ling Wang and institutional funding to Ah-Ng Tony Kong. The authors thank all the members in Dr. Kong’s lab for their helpful discussion and preparation of this manuscript.

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Authors and Affiliations

  1. Department of Clinical Pharmacy and Pharmacy Administration, West China School of Pharmacy, Sichuan University, 610041, Chengdu, Sichuan, China

    Ling Wang

  2. Center for Cancer Prevention Research, Department of Pharmaceutics, Rutgers, The State University of New Jersey, Piscataway, New Jersey, 08854-8020, USA

    Ling Wang, Chengyue Zhang, Yue Guo, Zheng-Yuan Su, Yuqing Yang, Limin Shu & Ah-Ng Tony Kong

  3. Department of Pharmaceutics & Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, New Jersey, 08854, USA

    Chengyue Zhang, Yue Guo, Zheng-Yuan Su, Yuqing Yang, Limin Shu & Ah-Ng Tony Kong

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  1. Ling Wang
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  2. Chengyue Zhang
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  3. Yue Guo
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  4. Zheng-Yuan Su
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  7. Ah-Ng Tony Kong
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Correspondence to Ling Wang or Ah-Ng Tony Kong.

Additional information

Guest Editors: Ah-Ng Tony Kong and Chi Chen

Ling Wang and Chengyue Zhang contributed equally to this work.

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Wang, L., Zhang, C., Guo, Y. et al. Blocking of JB6 Cell Transformation by Tanshinone IIA: Epigenetic Reactivation of Nrf2 Antioxidative Stress Pathway. AAPS J 16, 1214–1225 (2014). https://doi.org/10.1208/s12248-014-9666-8

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  • DOI: https://doi.org/10.1208/s12248-014-9666-8

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