Antrodin C Isolated from Antrodia Cinnamomea Induced Apoptosis through ROS/AKT/ERK/P38 Signaling Pathway and Epigenetic Histone Acetylation of TNFα in Colorectal Cancer Cells

Yung-Yu Hsieh; Ko-Chao Lee; Kung-Chuan Cheng; Kam-Fai Lee; Ya-Ling Yang; Hsin-Tung Chu; Ting-Wei Lin; Chin-Chu Chen; Meng-Chiao Hsieh; Cheng-Yi Huang; Hsing-Chun Kuo; Chih-Chuan Teng

doi:10.3390/antiox12030764

Antrodin C Isolated from Antrodia Cinnamomea Induced Apoptosis through ROS/AKT/ERK/P38 Signaling Pathway and Epigenetic Histone Acetylation of TNFα in Colorectal Cancer Cells

Antioxidants (Basel). 2023 Mar 21;12(3):764. doi: 10.3390/antiox12030764.

Authors

Yung-Yu Hsieh^{1

2}, Ko-Chao Lee^{3

4}, Kung-Chuan Cheng³, Kam-Fai Lee⁵, Ya-Ling Yang⁶, Hsin-Tung Chu⁷, Ting-Wei Lin⁷, Chin-Chu Chen⁷, Meng-Chiao Hsieh⁸, Cheng-Yi Huang⁸, Hsing-Chun Kuo^{9

10

11

12}, Chih-Chuan Teng^{9

10}

Affiliations

¹ Division of Gastroenterology and Hepatology, Department of Internal Medicine, Chang Gung Memorial Hospital, Chiayi 613016, Taiwan.
² College of Medicine, Chang Gung University, Taoyuan 333323, Taiwan.
³ Division of Colorectal Surgery, Department of Surgery, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Kaohsiung 833401, Taiwan.
⁴ College of Medicine, Chang Gung University, Kaohsiung 833401, Taiwan.
⁵ Department of Pathology, Chang Gung Memorial Hospital, Chiayi 613016, Taiwan.
⁶ Department of Anesthesiology, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung 833401, Taiwan.
⁷ Biotech Research Institute, Grap King Bio Ltd., Taoyuan 325002, Taiwan.
⁸ Division of Colon and Rectal Surgery, Department of Surgery, Chang Gung Memorial Hospital, Chiayi 613016, Taiwan.
⁹ Department of Nursing, Division of Basic Medical Sciences, Chang Gung University of Science and Technology, Chiayi 613016, Taiwan.
¹⁰ Research Fellow, Chang Gung Memorial Hospital, Chiayi 613016, Taiwan.
¹¹ Research Center for Food and Cosmetic Safety, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan 333324, Taiwan.
¹² Chronic Diseases and Health Promotion Research Center, Chang Gung University of Science and Technology, Chiayi 613016, Taiwan.

Abstract

Background: Antrodin C, a maleimide derivative compound isolated from the ethanol extract of the mycelium of Antrodia cinnamomea, is an endemic fungus of Taiwan and a potential chemoprotective agent. However, the molecular mechanisms underlying the mode of action of antrodin C on cancer cells, especially in human colorectal cancer (CRC), remain unclear.

Methods: The cell death and ROS of the antrodin-C-treated HCT-116 cells were measured by annexin V-FITC/propidium iodide staining, DCFDA, and Fluo-3 fluorescence staining assays. Moreover, signaling molecules regulating TNFα cell death pathways and ROS/AKT/ERK/P38 pathways were also detected in cells treated with antrodin C by Western blotting and chromatin immunoprecipitation. The effects of antrodin C were determined in HCT-116 cell xenograft animal models in terms of tumor volumes and histopathological evaluation.

Results: Treatment with antrodin C triggered the activation of extrinsic apoptosis pathways (TNFα, Bax, caspase-3, and -9), and also suppressed the expression of anti-apoptotic molecules Bcl-2 in HCT-116 cells in a time-dependent manner. Antrodin C also decreased cell proliferation and growth through the inactivation of cyclin D1/cyclin for the arrest of the cell cycle at the G1 phase. The activation of the ROS/AKT/ERK/P38 pathways was involved in antrodin-C-induced transcriptional activation, which implicates the role of the histone H3K9K14ac (Acetyl Lys9/Lys14) of the TNFα promoters. Immunohistochemical analyses revealed that antrodin C treatment significantly induced TNFα levels, whereas it decreased the levels of PCNA, cyclin D1, cyclin E, and MMP-9 in an in vivo xenograft mouse model. Thus, antrodin C induces cell apoptosis via the activation of the ROS/AKT/ERK/P38 signaling modules, indicating a new mechanism for antrodin C to treat CRC in vitro and in vivo.

Keywords: H3K9K14ac; TNFα; antrodin C; apoptosis; colorectal cancer.

Grants and funding

Funding for this study was provided in part by the research grants BMRPD42, CLRPG8L0063, CMRPF6L0061, CMRPF6L0062, CMRPF6L0063, and ZRRPF6M0011 as well as CMRPF6M0071, CMRPF6M0072, CMRPF6M0073 from Chang Gung Memorial Hospital, Chiayi, Taiwan, and Chang Gung University of Science and Technology, Chiayi Campus, Taiwan. This work was supported by the research grants MOST 110-2320-B-255-005-MY3 from the Ministry of Science and Technology, Taiwan.