PGV-1 Promotes Apoptosis via Mitotic Arrest and Senescence in Polyploid Giant Cancer Cells of Hepatocellular Carcinoma JHH4

INTRODUCTION: Senescent cells release a senescence-associated secretory phenotype, promoting Polyploid Giant Cancer Cells (PGCCs) emergence, fostering tumor heterogeneity and resistance. Pentagamavunone-1 (PGV-1) emerges as a promising agent inducing senescence and prometaphase arrest, resulting in permanent cytotoxicity. This study investigates the role of PGV-1 in dysregulating mitosis through the modulation of PGCCs and senescence in low MYCN-expressing hepatocellular carcinoma (HCC) cells, JHH4.
METHODS: To assess the physiological effects of PGV-1, we employed several in vitro tests, including the MTT assay, cell cycle assay, May-Grünwald-Giemsa staining, Senescence Associated–β–galactosidase (SA-β-Gal) assay, and apoptosis assay. The protein level of apoptosis regulatory protein was evaluated using western blot analysis. Interaction of PGV-1 toward protein played a role in PGCCs formation was simulated by molecular docking and molecular dynamic.
RESULTS: The cytotoxic assay revealed that PGV inhibited the proliferation of JHH4 liver cancer cells permanently. Inhibition proliferation by PGV-1 was associated with the modulation of G2/M phase particularly mitotic arrest and formation of PGCCs. The SA-β-Gal verified that PGV-1 induced senescence in cells with PGCCs formation. Apoptotic mechanisms were validated via annexin V staining and increased level of cleaved PARP. Molecular docking and molecular dynamic simulations suggested that PGV-1 could interfere with the conformation of chromosomal passenger complex (CPC) protein, particularly disrupting essential interactions within INCENP, Survivin, and Borealin. These interactions could cause conformational destabilization of CPC, leading to mitotic failure.
DISCUSSION AND CONCLUSION: Clinically, the ability of PGV-1 to target key mitotic regulators and induce a dual mechanism of senescence and apoptosis in HCC suggest its promise as a chemotherapeutic agent. By exploiting the vulnerabilities of mitotic machinery and PGCC, PGV-1 offers a new wave for treating aggressive liver cancers with high recurrence rates