BACKGROUND
Triple-negative breast cancer (TNBC) is a clinically challenging disease subtype of breast cancer, with therapeutic outcomes remaining unsatisfactory due to its highly invasive potential. Ferroptosis, a newly identified type of programmed cell death, is mediated by the iron-dependent accumulation of lipid peroxides, presenting a potential therapeutic avenue for the treatment of TNBC. Calycosin (Cal) has demonstrated antitumor effects across various tumors; however, its therapeutic potential based on the ferroptosis pathway for TNBC treatment and the associated mechanisms remains unclear.
METHODS
To achieve this, the cell counting kit-8 assay, flow cytometry assay, wound healing assay, colony formation assay, EdU assay, and xenograft mouse models were performed. Mechanisms were investigated using transcriptomic sequencing, quantitative reverse transcription polymerase chain reaction, and Western blot in vitro and in vivo.
RESULTS
Our findings indicated that Cal significantly reduces the viability of TNBC cells, primarily manifested as suppression of growth, migration, and colony formation through the induction of ferroptosis. Additionally, Cal impairs mitochondrial function in TNBC cells by reducing mitochondrial membrane potential and elevating ferroptosis-related markers, including MDA and 4-HNE accumulation, reactive oxygen species formation, and GSH depletion. Mechanistically, Cal administration may reduce GPX4 levels through the OTUD6B-mediated ubiquitination pathway, thereby enhancing the ubiquitination and degradation of the GPX4 protein. The decreased OTUD6B and GPX4 expression significantly inhibited TNBC growth both in vivo and in vitro, resulting in ferroptosis induction in TNBC cells.
CONCLUSION
This is the first study to demonstrate that Cal exerts anti-TNBC activity through a novel OTUD6B/GPX4-dependent ferroptosis regulatory axis. These findings highlight that Cal warrants further investigation as a potential candidate, and the OTUD6B/GPX4 signaling pathway may represent a significant target for TNBC treatment.
Xue Mao, Yin Lin, Zhifeng Gu et al.· Biochemical and Biophysical...· 0 citations
Aplastic anemia (AA) is a severe hematological disorder caused by hyperactivated T cell-mediated hematopoietic failure. It is characterized by hematopoietic stem cell deficiency and hypocellular hematopoiesis in the bone marrow (BM). Ferroptosis, a specific type of programmed cell death, is defined by iron-dependent lipid peroxidation, which has garnered the attention of researchers due to its unique role and biological importance in various diseases. However, the correlation between immunological imbalance-induced ferroptosis and the mortality of hematopoietic stem cells within the BM microenvironment in AA is unclear. Resveratrol (RSV) is crucial in activating NRF2, hence influencing the onset and progression of diseases by regulating ferroptosis. This study aimed to investigate the roles and molecular mechanisms of RSV in the hematopoietic recovery of AA regarding ferroptosis. We measured some biomarkers in AA mice and IFN-gamma-treated 32D cells, representing AA syndromes and ferroptosis features. Furthermore, in RSV-treated 32D AA cells, cell activity, cell apoptosis, mitochondrial membrane potential, mitochondrial membrane permeability, and intracellular levels of MDA, ferrous iron, 4-HNE, GSH, ROS, and lipid peroxidation were assessed. Subsequently, additional in vitro and in vivo experiments were performed to investigate the mechanisms by which RSV effectively regulates NRF2 stability, further inhibiting ferroptosis in AA. We demonstrated that ferroptosis contributes to the occurrence and development of AA disease. RSV dose-dependently inhibits ferroptosis in 32D AA cells by targeting GPX4 expression, enhancing the cell activity of hematopoietic BM cells. Mechanically, RSV significantly increases NRF2 phosphorylation through the PI3K/AKT/mTOR signaling pathway, which maintains NRF2 stability to promote the GPX4 metabolic pathway in AA. Protein levels of p-AKT, p-mTOR, p-NRF2, and GPX4 were markedly increased in BM cells in RSV-treated AA mice, resulting in the suppression of AA development. Our data demonstrate that RSV effectively inhibits ferroptosis in AA or IFN-gamma-treated 32D cells by targeting the PI3K/AKT/mTOR signaling pathway, which enhances NRF2-mediated GPX4 transcription, thereby ameliorating AA symptoms in vitro and in vivo. This study concludes that RSV is a potential therapeutic drug for ferroptosis-related AA treatment.
Erhao Zhang, Jianan Cui, Xinran Zhu et al.· Chinese Medicine· 0 citations
