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Targeting Nrf2 with wogonin overcomes cisplatin resistance in head and neck cancer

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Abstract

A principal limitation to the clinical use of cisplatin is the high incidence of chemoresistance to this drug. Combination treatments with other drugs may help to circumvent this problem. Wogonin, one of the major natural flavonoids, is known to reverse multidrug resistance in several types of cancers. We investigated the ability of wogonin to overcome cisplatin resistance in head and neck cancer (HNC) cells and further clarified its molecular mechanisms of action. Two cisplatin-resistant HNC cell lines (AMC-HN4R and -HN9R) and their parental and other human HNC cell lines were used. The effects of wogonin, either alone or in combination with cisplatin, were assessed in HNC cells and normal cells using cell cycle and death assays and by measuring cell viability, reactive oxygen species (ROS) production, and protein expression, and in tumor xenograft mouse models. Wogonin selectively killed HNC cells but spared normal cells. It inhibited nuclear factor erythroid 2-related factor 2 and glutathione S-transferase P in cisplatin-resistant HNC cells, resulting in increased ROS accumulation in HNC cells, an effect that could be blocked by the antioxidant N-acetyl-l-cysteine. Wogonin also induced selective cell death by targeting the antioxidant defense mechanisms enhanced in the resistant HNC cells and activating cell death pathways involving PUMA and PARP. Hence, wogonin significantly sensitized resistant HNC cells to cisplatin both in vitro and in vivo. Wogonin is a promising anticancer candidate that induces ROS accumulation and selective cytotoxicity in HNC cells and can help to overcome cisplatin-resistance in this cancer.

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Abbreviations

HNC:

Head and neck cancer

Nrf2:

Nf-E2-related factor 2

ARE:

Antioxidant response element

ROS:

Reactive oxygen species

NAC:

N-acetyl-l-cysteine

GSH:

Glutathione

GSSG:

Glutathione disulfide

DCF-DA:

2′,7′-Dichlorofluorescein diacetate

PARP:

Poly(ADP-ribose) polymerase

JNK:

c-Jun N-terminal kinase

GSTP1:

Glutathione S-transferase pi 1

siRNA:

Short interfering RNA

TUNEL:

Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling.

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Acknowledgments

This study was supported by a Grant (No. 2015R1A2A1A15054540) from Basic Science Research Program through the National Research Foundation of Korea (NRF), Ministry of Science, ICT, and Future Planning, and a Grant (No. HI15C2920) from the Korean Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), Ministry of Health & Welfare, Seoul, Republic of Korea (J.-L. Roh).

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Authors and Affiliations

  1. Department of Otolaryngology, Asan Medical Center, University of Ulsan College of Medicine, 88 Olympic-ro 43-gil, Songpa-gu, Seoul, 05505, Republic of Korea

    Eun Hye Kim, Hyejin Jang, Daiha Shin, Seung Ho Baek & Jong-Lyel Roh

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  1. Eun Hye Kim
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Correspondence to Jong-Lyel Roh.

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Kim, E.H., Jang, H., Shin, D. et al. Targeting Nrf2 with wogonin overcomes cisplatin resistance in head and neck cancer. Apoptosis 21, 1265–1278 (2016). https://doi.org/10.1007/s10495-016-1284-8

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