Research Paper Volume 13, Issue 12 pp 15964—15989
Antrodia salmonea induces apoptosis and enhances cytoprotective autophagy in colon cancer cells
- 1 Institute of Nutrition, College of Health Care, China Medical University, Taichung 40402, Taiwan
- 2 Department of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, College of Chinese Medicine, China Medical University, Taichung 40402, Taiwan
- 3 Department of Cosmeceutics, College of Pharmacy, China Medical University, Taichung 40402, Taiwan
- 4 Department of Health and Nutrition Biotechnology, Asia University, Taichung 41354, Taiwan
- 5 Chinese Medicine Research Center, China Medical University, Taichung 40402, Taiwan
Received: February 3, 2021 Accepted: April 5, 2021 Published: May 24, 2021
https://doi.org/10.18632/aging.203019How to Cite
Copyright: © 2021 Yang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
A traditional Chinese medicinal fungus, Antrodia salmonea (AS), with antioxidant properties is familiar in Taiwan but anti-cancer activity of AS in human colon cancer is ambiguous. Hence, we explored the anti-cancer activity of AS in colon cancer cells. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay revealed that AS showed a remarkable effect on cell viability in colon cancer cells; SW620, HCT116, and HT29. Annexin V/propidium iodide (PI) stained cells indicated that AS induced both early/late apoptosis in SW620 cells. Additionally, cells treated with AS induced caspase-3 activation, poly (ADP-ribose) polymerase (PARP) cleavage, mitochondrial dysfunction, and Bcl-2 associated X (Bax)/B-cell lymphoma (Bcl-2) dysregulation. Microtubule- associated protein 1A/1B-light chain 3B (LC3-II) accumulation, sequestosome 1 (p62/SQSTM1) activation, autophagy related 4B cysteine peptidase (ATG4B) inactivation, acidic vesicular organelles (AVOs) formation, and Beclin-1/Bcl-2 dysregulation revealed that AS-induced autophagy. Interestingly, cells pretreated with 3-methyladenine (3-MA) strengthened AS-induced caspase-3/apoptosis. Suppression of apoptosis by z-Val-Ala-Asp fluoromethyl ketone (Z-VAD-FMK) did not however block AS-induced autophagy, suggesting that autophagy was not attenuated by the AS-induced apoptosis. Application of N-acetylcysteine (NAC) prevented AS-induced cell death, caspase-3 activation, LC3-II accumulation, and AVOs formation, indicating that AS-induced apoptosis and autophagy was mediated by reactive oxygen species (ROS). Furthermore, AS-induced cytoprotective autophagy and apoptosis through extracellular signal-regulated kinase (ERK) signaling cascades. Moreover, in vivo data disclosed that AS inhibited colitis-associated tumorigenesis in azoxymethane (AOM)-dextran sodium sulphate (DSS)-treated mice. For the first time, we report the anti-cancer properties of this potentially advantageous mushroom for the treatment of human colon cancer.
Abbreviations
AKT: serine/threonine kinase; AO: acridine orange; AOM: Azoxymethane; ATG4B: autophagy related 4B cysteine peptidase; AS: Antrodia salmonea; AVOs: acidic vesicular organelles; BSA: bovine serum albumin; CAC: colitis-associated cancer; CQ: chloroquine; DCFH2-DA: 2′-7′ dihydrofluorescein diacetate; DSS: dextran sulfate sodium; ERK1/2: extracellular signal regulated kinase 1/2; FBS: fetal bovine serum; FITC: fluorescein isothiocyanate; HE: hematoxylin and eosin; LC3: microtubule-associated protein light chain 3; mTOR: mammalian target of rapamycin MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylhtetrazolium bromide; 3-MA: 3-methyladenine; NAC: N-acetylcysteine; NFκB: nuclear factor kappa B; PARP: poly ADP ribose polymerase; PI: propidium iodide; p62/SQSTM1: sequestosome 1; ROS: reactive oxygen species; Z-VAD-FMK: benzyloxycarbonyl-Val-Ala-Asp (OMe) fluoromethylketone.