Figure 2 from Targeting Cancer-Associated PCNA with AOH1996 Induces Mitotic Catastrophe and Enhances Cisplatin Therapy in Cervical Cancer

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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.

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