Bifunctional mechanisms of autophagy and apoptosis regulations in melanoma from Bacillus subtilis natto fermentation extract

Hsin-Yu Chou; Li-Heng Liu; Chung-Yi Chen; I-Fan Lin; Daoud Ali; Alan Yueh-Luen Lee; Hui-Min David Wang

doi:10.1016/j.fct.2021.112020

Bifunctional mechanisms of autophagy and apoptosis regulations in melanoma from Bacillus subtilis natto fermentation extract

Food Chem Toxicol. 2021 Apr:150:112020. doi: 10.1016/j.fct.2021.112020. Epub 2021 Jan 26.

Authors

Hsin-Yu Chou¹, Li-Heng Liu², Chung-Yi Chen³, I-Fan Lin⁴, Daoud Ali⁵, Alan Yueh-Luen Lee⁶, Hui-Min David Wang⁷

Affiliations

¹ Program in Tissue Engineering and Regenerative Medicine, National Chung Hsing University, Taichung 402, Taiwan.
² Institute of Molecular Medicine, College of Medicine, National Taiwan University, Taipei 10617, Taiwan.
³ School of Medical and Health Sciences, Fooyin University, Kaohsiung 831, Taiwan.
⁴ Challenge Bioproducts Co., Ltd., Yunlin County 640, Taiwan.
⁵ Department of Zoology, College of Science, King Saud University BOX 2455, Saudi Arabia Riyadh 11451, Saudi Arabia.
⁶ Program in Tissue Engineering and Regenerative Medicine, National Chung Hsing University, Taichung 402, Taiwan; National Institute of Cancer Research, National Health Research Institutes, Zhunan, Miaoli 35053, Taiwan; Department of Biotechnology, College of Life Science, Kaohsiung Medical University, Kaohsiung 80708, Taiwan. Electronic address: alanylee@nhri.edu.tw.
⁷ Program in Tissue Engineering and Regenerative Medicine, National Chung Hsing University, Taichung 402, Taiwan; Graduate Institute of Biomedical Engineering, National Chung Hsing University, Taichung City 402, Taiwan; Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung City 807, Taiwan; Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung City 404, Taiwan. Electronic address: davidw@dragon.nchu.edu.tw.

PMID: 33513408
DOI: 10.1016/j.fct.2021.112020

Abstract

Melanoma is one of the most dangerous malignant epidermal cancers. Natto freeze-drying extract (NFDE) and natto water extract (NWE) were isolated from natto, soybeans fermented by Bacillus subtilis natto, which were assessed as potential anti-melanoma agents. Cell cytotoxicity assays revealed significant anti-melanoma effects of NFDE and NWE in a dose-dependent manner, and exhibited low influences on normal skin cells, including Hs68, HaCaT and adipose tissue-derived stem cells (ADSCs), respectively. Through a flow cytometer assay and autophagy acridine orange staining, the cellular death phenomenon shifted from autophagy to apoptosis with the increased dosages. Reactive oxygen species (ROS) were enhanced using DCFDA (2,7-dichlorodihydrofluorescein diacetate) staining when melanoma cells were treated with the extract. NFDE and NWE treatments increase the oxidative stress of cancer cells and cause apoptosis by inhibiting AMP-activated protein kinase (AMPK). NFDE and NWE were considered to play a critical role in cell death through ROS adjustment, autophagy regulation and apoptosis promotion.

Keywords: Apoptosis; Autophagy; Bacillus subtilis natto Extract; Melanoma; Oxidative stress.

MeSH terms

Animals
Apoptosis / drug effects*
Autophagy / drug effects*
Bacillus subtilis / chemistry
Bacillus subtilis / physiology*
Cell Line
Cell Survival / drug effects
Fermentation
Fibroblasts
Gene Expression Regulation, Neoplastic / drug effects
Glycine max / metabolism
Humans
Japan
Melanoma / drug therapy*
Mice
Soy Foods