Nugraheni Nadzifa, Lestari Beni, Meiyanto Edy, Utomo Rohmad Yudi
Universitas Gadjah Mada, Graduate School of Biotechnology, Yogyakarta, Indonesia.
Universitas Gadjah Mada, Faculty of Medicine, Department of Pharmacology and Therapy, Division of Public Health and Nursing, Yogyakarta, Indonesia.
Turk J Pharm Sci. 2025 Aug 1;22(3):170-177. doi: 10.4274/tjps.galenos.2025.46837.
Senescent cells release a senescence-associated secretory phenotype, promoting polyploid giant cancer cells (PGCCs) to emerge, 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 was aimed to investigate the effect of PGV-1 in dysregulating mitosis through the modulation of PGCCs and senescence in low MYCN-expressing hepatocellular carcinoma (HCC) cells JHH4.
To assess the physiological effects of PGV-1, several tests were done including the MTT assay, cell cycle assay, May-Grünwald-Giemsa staining, senescence associated-β-galactosidase (SA-β-Gal) assay, and apoptosis assay. The protein levels of the apoptosis regulatory protein were evaluated using western blot analysis. The interaction of PGV-1 toward the protein that plays a role in PGCCs formation was simulated by molecular docking and molecular dynamics (MD).
The cytotoxic assay revealed that PGV-1 inhibited the proliferation of JHH4 liver cancer cells permanently. Inhibition of cell 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 (<0.01), inducing PGCCs formation. Apoptotic mechanisms were validated Annexin V staining, which showed the level of cleaved poly (ADP-ribose) polymerase (<0.001). Molecular docking and MD simulations suggested that PGV-1 could interfere with the conformation of the chromosomal passenger complex (CPC) protein, particularly disrupting essential interactions within the inner centromere protein, Survivin, and Borealin.
PGV-1 induced strong cytotoxicity in HCC cells by disrupting mitosis leading to PGCC formation, senescence, and subsequent apoptotic cell death.
衰老细胞释放衰老相关分泌表型，促使多倍体巨癌细胞（PGCCs）出现，促进肿瘤异质性和耐药性。五环戊烯酮-1（PGV-1）成为一种有前景的诱导衰老和前中期阻滞的药物，导致永久性细胞毒性。本研究旨在探讨PGV-1通过调节低MYCN表达的肝癌（HCC）细胞JHH4中的PGCCs和衰老来失调有丝分裂的作用。
为评估PGV-1的生理作用，进行了多项试验，包括MTT试验、细胞周期试验、May-Grünwald-Giemsa染色、衰老相关β-半乳糖苷酶（SA-β-Gal）试验和凋亡试验。使用蛋白质印迹分析评估凋亡调节蛋白的蛋白质水平。通过分子对接和分子动力学（MD）模拟PGV-1与在PGCCs形成中起作用的蛋白质的相互作用。
细胞毒性试验表明，PGV-1可永久抑制JHH4肝癌细胞的增殖。PGV-1对细胞增殖的抑制与G2/M期的调节有关，特别是有丝分裂阻滞和PGCCs的形成。SA-β-Gal证实PGV-1可诱导细胞衰老（<0.01），诱导PGCCs形成。通过膜联蛋白V染色验证了凋亡机制，其显示裂解的聚（ADP-核糖）聚合酶水平（<0.001）。分子对接和MD模拟表明，PGV-1可干扰染色体乘客复合体（CPC）蛋白的构象，特别是破坏着丝粒内蛋白、生存素和勃林蛋白之间的关键相互作用。
PGV-1通过破坏有丝分裂导致PGCC形成、衰老及随后的凋亡性细胞死亡，从而在HCC细胞中诱导强烈的细胞毒性。
