G-4 inhibits triple negative breast cancer by inducing cell apoptosis and promoting LCN2-dependent ferroptosis

Area of Science:

• Oncology
• Molecular Biology
• Biochemistry

Background:

• Triple-negative breast cancer (TNBC) remains a challenging subtype with limited targeted therapies.
• Rocovitine derivatives have shown potential as cyclin-dependent kinase inhibitors.
• Understanding novel therapeutic mechanisms is crucial for advancing TNBC treatment.

Purpose of the Study:

• To investigate the antitumor activity, mechanism of action, and potential targets of compound G-4 in TNBC.
• To elucidate the role of G-4 in cell cycle regulation, apoptosis, and ferroptosis.
• To explore the relationship between G-4, EGFR/AKT/MAPK pathways, and LCN2 expression.

Main Methods:

• Flow cytometry and immunoblotting to assess cell cycle arrest and apoptosis.
• RNA sequencing (RNA-seq) to identify differentially expressed genes.
• siRNA transfection to create a LCN2 knockdown model.
• Measurement of reactive oxygen species (ROS), lipid ROS, iron, MDA, SOD, and GSH levels.

Main Results:

• G-4 induced S phase arrest and mitochondrial apoptosis by inhibiting EGFR, AKT, and MAPK pathways.
• G-4 triggered iron-mutagenesis, elevated ROS and lipid ROS, and down-regulated LCN2.
• LCN2 knockdown significantly reduced G-4 sensitivity and ferroptosis markers.

Conclusions:

• Compound G-4 exhibits potent antitumor activity against TNBC by inducing apoptosis via EGFR pathway inhibition.
• G-4 promotes LCN2-dependent ferroptosis, presenting a novel therapeutic avenue.
• G-4 represents a promising candidate for TNBC treatment, targeting both apoptosis and ferroptosis pathways.