Figure 2.

AOH1996 selectively induces apoptosis in cervical cancer (CaCx) cells. A, Dose–response curves of AOH1996 in primary HFKs, HeLa (HPV18⁺), CaSki (HPV16⁺), C33A (HPV⁻), and HeLa cells overexpressing POLη following 72 hours of treatment, assessed via MTT assay. HFKs show resistance, whereas all CaCx cell lines are sensitive to nanomolar concentrations. B, Calculated GI₅₀ values confirm significantly greater sensitivity of CaCx lines to AOH1996 compared with HFKs. C, Representative Annexin V/PI flow cytometry plots after 48 hours of AOH1996 treatment (1 μmol/L) show live (Annexin V⁻/PI⁻), apoptotic (Annexin V⁺/PI⁻), and dead (Annexin V⁺/PI⁺ or Annexin V⁻/PI⁺) populations. D, Quantification of flow cytometry data indicates robust induction of apoptosis and cell death in all CaCx lines, with negligible effects in HFKs. E, CaSki organoid cultures treated with 3 μmol/L AOH1996 show increased cell death and reduced proliferation. Hematoxylin and eosin (H&E) staining highlights stratified epithelial structure and a dying cell zone upon AOH1996 or cisplatin treatment. F, Quantification of H&E-stained rafts by a pathologist. G, TUNEL (green) and γH2AX (red) staining indicate increased apoptosis and DNA damage in response to AOH1996. Graphs show intensity quantification and quantification of relative thickness. Data represent the mean ± SEM from at least three replicates. Statistical analysis by one-way ANOVA. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.