Digoxigenin activates autophagy in hepatocellular carcinoma cells by regulating the PI3K/AKT/mTOR pathway

Area of Science:

• Oncology
• Pharmacology
• Biochemistry

Background:

• Hepatocellular carcinoma (HCC) is a highly malignant tumor with limited effective treatments.
• Current targeted combination immunotherapy for HCC faces challenges with adverse effects and low response rates.
• Traditional Chinese medicine offers low-toxicity, cost-effective anticancer agents.

Purpose of the Study:

• To identify novel anti-HCC drugs using high-throughput phenotypic screening.
• To investigate the therapeutic potential and mechanism of action of digoxigenin (DIG) against HCC.
• To explore DIG's effects on the PI3K/AKT/mTOR signaling pathway and autophagy.

Main Methods:

• High-throughput phenotypic screening of 1,444 bioactive compounds.
• In vitro validation using CCK8, lactate dehydrogenase, and colony formation assays.
• Mechanistic studies including transmission electron microscopy, western blotting, immunofluorescence, network pharmacology, and molecular docking.
• In vivo assessment using subcutaneous xenograft tumor models.

Main Results:

• Digoxigenin (DIG) significantly impeded HCC cell progression and proliferation in vitro.
• DIG was demonstrated to inhibit HCC cell proliferation by inducing autophagy.
• Network pharmacology and molecular docking indicated DIG targets the PI3K/AKT/mTOR pathway, confirmed by comparative treatments.
• In vivo studies showed DIG effectively halted subcutaneous xenograft tumor growth.

Conclusions:

• Digoxigenin (DIG) is a promising therapeutic agent for hepatocellular carcinoma (HCC).
• DIG exerts its anti-HCC effects by modulating the PI3K/AKT/mTOR pathway to induce autophagy.
• Phenotypic screening accelerates the discovery of novel HCC therapies targeting key molecular pathways.