Resibufogenin inhibited colorectal cancer cell growth and tumorigenesis through triggering ferroptosis and ROS production mediated by GPX4 inactivation.

Abstract

Resibufogenin (RB) has been used for cancer treatment, but the underlying mechanisms are still unclear. This study aimed to investigate the effects of RB treatment on colorectal cancer (CRC) cells, and to determine the underlying mechanisms. The cell counting kit-8 assay was used to determine cell viability. Cell morphology was observed under light microscopy, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay was employed to detect cell apoptosis. Intracellular ferrous iron (Fe2+ ), malondialdehyde (MDA), glutathione (GSH), and reactive oxygen species levels were detected by using commercial iron assay kit, MDA assay kit, GSH assay kit, and 2,7-dichlorodihydrofluorescein diacetate probes, respectively. The protein expressions were determined by Western blot and immunohistochemistry. RB inhibited cell viability in the CRC cell lines (HT29 and SW480) in a dose- and time-dependent manner, and caused cytotoxicity to the normal colonic epithelial cell line (NCM460) at high dose. Similarly, RB induced morphological changes in CRC cells from normal to round shape, and promoted cell death. Of note, RB triggered oxidative stress and ferroptotic cell death in CRC cells, and only ferroptosis inhibitors (deferoxamine and ferrostatin-1), instead of inhibitors for other types of cell death (apoptosis, autophagy, and necroptosis), reversed the inhibitory effects of RB on CRC cell proliferation. Furthermore, glutathione peroxidase 4 (GPX4) was inactivated by RB treatment, and overexpression of GPX4 alleviated RB-induced oxidative cell death in CRC cells. Consistently, the in vivo experiments validated that RB also triggered oxidative stress, and inhibited CRC cells growth and tumorigenicity in mice models. RB can inhibit CRC cells growth and tumorigenesis by triggering ferroptotic cell death in a GPX4 inactivation-dependent manner.

• Authors: Lian-Dong Shen, Wen-Hai Qi, Jiang-Jiang Bai, Chun-Yi Zuo, Dong-Lin Bai, Wei-Dong Gao, Xin-Ling Zong, Ting-Ting Hao, Yan Ma, Guang-Cai Cao
• Journal: Anatomical record (Hoboken, N.J. : 2007) (Anat Rec (Hoboken)) Vol. 304 Issue 2 Pg. 313-322 (02 2021) ISSN: 1932-8494 [Electronic] United States
• PMID: 31961485 (Publication Type: Journal Article, Retracted Publication)
• Copyright: © 2020 American Association for Anatomy.

Chemical References

• Bufanolides
• Reactive Oxygen Species
• bufogenin
• Phospholipid Hydroperoxide Glutathione Peroxidase

Topics

• Bufanolides (pharmacology)
• Carcinogenesis (drug effects)
• Cell Death (drug effects)
• Cell Line, Tumor
• Cell Proliferation (drug effects)
• Cell Shape (drug effects)
• Cell Survival (drug effects)
• Colorectal Neoplasms (metabolism, pathology)
• Ferroptosis (drug effects)
• Humans
• Oxidative Stress (drug effects)
• Phospholipid Hydroperoxide Glutathione Peroxidase (metabolism)
• Reactive Oxygen Species (metabolism)