A novel NRF2/ARE inhibitor gossypol induces cytotoxicity and sensitizes chemotherapy responses in chemo-refractory cancer cells

Ya-Chu Tang, Graduate Program of Biotechnology in Medicine, Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu, Taiwan
Hsin-Huei Chang, Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, Miaoli, Taiwan
Huang-Hui Chen, Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, Miaoli, Taiwan
Jau-Ying Yao, Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, Miaoli, Taiwan
Yu-Tsen Chen, National Institute of Cancer Research, National Health Research Institutes, Miaoli, Taiwan
Yung-Jen Chuang, Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu, Taiwan
Jang-Yang Chang, Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, Miaoli, Taiwan
Ching-Chuan Kuo, Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, Miaoli, TaiwanFollow

Abstract

NRF2/ARE signaling pathway is a principal regulator of cellular redox homoeostasis. The stress-induced transcription factor, NRF2, can shield cells from the oxidative damages via binding to the consensus antioxidant-responsive element (ARE) and driving several cyto-protective genes expression. Increasing evidence indicated that aberrant activation of NRF2 in malignant cells may support their survival through various pathways to detoxify chemotherapy drugs, attenuate drug-induced oxidative stress, or induce drug efflux, all of which are crucial in developing drug resistance. Accordingly, NRF2 is a potential drug target for improving the effectiveness of chemotherapy and to reverse drug resistance in cancer cells. A stable ARE-driven reporter human head and neck squamous cell carcinoma (HNSCC) cell line, HSC3-ARE9, was established and utilized to screen novel NRF2 inhibitors from a compound library. The cotton plant derived phenolic aldehyde-gossypol was selected for further analyses. The effects of gossypol in cancer cells were determined by western blotting, RT-qPCR, clonogenic assay, and cell viability assays. The gossypol-responsive gene expression levels were assessed in the Oncomine database. The effects of gossypol on conferring chemo-sensitization were evaluated in etoposide-resistant and cisplatin-resistant cancer cells. Our study is the first to identify that gossypol is effective to reduce both basal and NRF2 activator tert-butylhydroquinone (t-BHQ)-induced ARE-luciferase activity. Gossypol diminishes NRF2 protein stability and thereby leads to the suppression of NRF2/ARE pathway, which resulted in decreasing the expression levels of NRF2 downstream genes in both time- and dose-dependent manners. Inhibition of NRF2 by gossypol significantly decreases cell viabilities in human cancer cells. In addition, we find that gossypol re-sensitizes topoisomerase II poison treatment in etoposide-resistant cancer cells via suppression of NRF2/ABCC1 axis. Moreover, gossypol suppresses NRF2-mediated G6PD expression thereby leads to induce synthetic lethality with cisplatin not only in parental cancer cells but also in cisplatin-resistant cancer cells. These findings suggest that gossypol is a novel NRF2/ARE inhibitor, and can be a potential adjuvant chemotherapeutic agent for treatment of chemo-refractory tumor.

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Recommended Citation

Tang, Ya-Chu; Chang, Hsin-Huei; Chen, Huang-Hui; Yao, Jau-Ying; Chen, Yu-Tsen; Chuang, Yung-Jen; Chang, Jang-Yang; and Kuo, Ching-Chuan (2021) "A novel NRF2/ARE inhibitor gossypol induces cytotoxicity and sensitizes chemotherapy responses in chemo-refractory cancer cells," Journal of Food and Drug Analysis: Vol. 29 : Iss. 4 , Article 7.
Available at: https://doi.org/10.38212/2224-6614.3376