Annurca apple polyphenol extract selectively kills MDA-MB-231 cells through ROS generation, sustained JNK activation and cell growth and survival inhibition

Elisa Martino,Nunzia D’Onofrio,Stefania D’Angelo,Marina Porcelli,Daniela Cristina Vuoso,Luigi Mele,Giovanna Cacciapuoti

doi:10.1038/s41598-019-49631-x

Elisa Martino, Nunzia D’Onofrio + Show 5 more

Open Access

https://doi.org/10.1038/s41598-019-49631-x

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Abstract

Polyphenols represent the most studied class of nutraceuticals that can be therapeutics for a large spectrum of diseases, including cancer. In this study, we investigated for the first time the antitumor activities of polyphenol extract from Annurca apple (APE) in MDA-MB-231 triple negative breast cancer cells, and we explored the underlying mechanisms. APE selectively inhibited MDA-MB-231 cell viability and caused G2/M phase arrest associated with p27 and phospho-cdc25C upregulation and with p21 downregulation. APE promoted reactive oxygen species (ROS) generation in MDA-MB-231 cells while it acted as antioxidant in non-tumorigenic MCF10A cells. We demonstrated that ROS generation represented the primary step of APE antitumor activity as pretreatment with antioxidant N-acetylcysteine (NAC) prevented APE-induced G2/M phase arrest, apoptosis, and autophagy. APE downregulated Dusp-1 and induced a significant increase in JNK/c-Jun phosphorylation that were both prevented by NAC. Moreover, downregulation of JNK by its specific inhibitor SP600125 significantly diminished the anticancer activity of APE indicating that ROS generation and sustained JNK activation represented the main underlying mechanism of APE-induced cell death. APE also inhibited AKT activation and downregulated several oncoproteins, such as NF-kB, c-myc, and β-catenin. In light of these results, APE may be an attractive candidate for drug development against triple negative breast cancer.

Highlights

Breast cancer represents the principal cause of cancer death among women in developed countries[1]
The present study showed that APE exhibited anticancer activities in MDA-MB-231 triple negative breast cancer cells and provided experimental evidences on the underlying mechanisms
We found that APE selectively and efficiently causes a time- and dose-dependent inhibition of MDA-MB-231 cell viability by a pro-oxidant cytotoxic effect while in non-tumorigenic MCF10A cells it exerted an antioxidant protective effect against the H2O2-induced oxidative stress

Summary

Introduction

Breast cancer represents the principal cause of cancer death among women in developed countries[1]. Adverse toxic side effects of chemotherapy during breast cancer treatment have shifted considerable focus towards anti-cancer natural compounds as a valuable source for new drug development. Dietary components and natural products enhance the efficacy of standard chemotherapy by overcoming drug resistance and by reducing toxicity and side-effects[4,5]. Polyphenols are good candidates to be used as inhibitors of tumor cells growth[7] for their potential to regulate the www.nature.com/scientificreports/. Www.nature.com/scientificreports activity of several gene targets involved in carcinogenesis, through both direct interaction and modulation of their expression. Polyphenol-induced ROS production could be an effective strategy for the selective killing of cancer cells[11]

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