Cristazine inhibits HPV-positive cervical cancer cells via inducing caspase-dependent apoptosis

Objective: To determine whether cristazine inhibits HPV-positive cervical cancer cells and to elucidate its underlying mechanisms. Methods: HeLa cells were treated with varying concentrations of cristazine, and cell viability, intracellular ROS, apoptosis, and apoptosis-related gene and protein expression were assessed using WST-1, DCF-DA flow cytometry, Annexin V-FITC/PI staining, quantitative RT-PCR, and Western blotting. Additionally, CaSki and SiHa cells were evaluated in viability assays, with selected ROS and apoptosis measurements performed in CaSki cells. Results: After 24 h of treatment, cristazine reduced HeLa cell viability with an IC 50 of 0.223 μM and increased intracellular ROS and apoptotic populations. Caspase inhibition substantially restored viability and reduced apoptosis. Cristazine also increased TP53, CDKN1A, BAX, BCL2 , and CASP3 expression, and reduced HPV18 E6/E7 expression. Additionally, Western blotting showed upregulated p53, p21 and Bax/Bcl-2 ratio, and increased cleavage of caspases-3, -8, -9, and -12 and PARP after cristazine treatment. DAPI staining showed chromatin condensation, while wound healing assays demonstrated reduced wound closure following cristazine treatment. Conclusions: Cristazine suppresses cervical cancer cell growth via coordinated ROS-associated, p53-driven, and caspase-dependent apoptotic signaling, highlighting its potential as a marine-derived anticancer candidate that warrants further investigation.