Bufalin Inhibits the PI3K/AKT Pathway by Targeting GTF3C4 to Impede Breast Cancer Progression

Area of Science:

• Oncology
• Pharmacology
• Molecular Biology

Background:

• Global breast cancer incidence is increasing, posing significant challenges.
• Existing treatments face issues like side effects and drug resistance.
• Bufalin, a bufadienolide, shows potential as an anti-cancer agent.

Purpose of the Study:

• To investigate the anti-cancer effects of bufalin on breast cancer.
• To identify the molecular target of bufalin in breast cancer.
• To elucidate the mechanism by which bufalin inhibits breast cancer progression.

Main Methods:

• Cell proliferation assays using MDA-MB-231 and MCF-7 cell lines.
• Limited Proteolysis-Mass Spectrometry (LP-MS) for target identification.
• Cell thermal shift assays (CETSA), drug affinity response target stability (DARTS), and surface plasmon resonance (SPR) for target validation.
• RNA sequencing and single-cell RNA sequencing for pathway analysis.
• In vivo and in vitro experiments involving bufalin treatment and GTF3C4 knockdown.

Main Results:

• Bufalin inhibited proliferation of breast cancer cells and protected mice against breast cancer.
• GTF3C4 was identified as a direct binding target of bufalin and is overexpressed in breast cancer.
• Bufalin suppressed the PI3K/AKT signaling pathway by reducing GTF3C4 levels.
• GTF3C4 knockdown mimicked bufalin's effects, inhibiting proliferation and promoting apoptosis.
• Bufalin modulated the tumor microenvironment by altering immune cell populations and cell-cell signaling.

Conclusions:

• Bufalin targets GTF3C4 to inhibit breast cancer progression.
• The mechanism involves suppression of the PI3K/AKT pathway and tumor microenvironment remodeling.
• Bufalin represents a potential therapeutic agent for breast cancer.