The neolignan Miliusin B inhibits cell migration and promotes apoptotic cell death in cervical cancer cell line HeLa.

Previously, we reported the discovery, chemical characterization, and preliminary anticancer activity of Miliusin B (MB), a natural neolignan from Miliusa sessilis, demonstrating cytotoxicity and selectivity against the human cervical cancer cell line HeLa. In the present study, we further investigated the underlying anticancer mechanisms of MB. Consistent with our earlier findings, MB exhibited sub-micromolar cytotoxicity toward HeLa cells with >60-fold selectivity relative to non-cancerous controls. Mechanistic analyses revealed that cytotoxic concentrations of MB (5 µM) induced G2/M cell-cycle arrest and activated apoptotic pathways, as evidenced by caspase-9, caspase-3 and PARP cleavage. In addition, MB significantly impaired HeLa cell migration at sub-toxic concentrations (≤ 0.5 µM). Its efficacy was further validated using a three-dimensional spheroid model, where MB suppressed spheroid growth at 0.5 and 5 µM. Finally, combinatorial assays with standard cisplatin (Cis)-based cervical cancer chemotherapeutics (Cis monotherapy and Cis doublet regimens) demonstrated that MB enhanced the anti-proliferative effects of all treatments tested. Collectively, these findings highlight MB as a promising selective cytotoxic and anti-migratory candidate for further development as a potential therapeutic agent against cervical cancer.