Figure 4.

AOH1996 induces mitotic arrest and mitotic death in cervical cancer cells. A, Representative images of nuclear phenotypes observed by live-cell imaging of HFK, HeLa, and HeLa POLη cells stably expressing GFP-LaminB1 (green) and mCherry-H2B (red). Cells were classified as normal, mitotic, irregular, or dead based on nuclear envelope integrity and chromatin morphology. Scale bars = 5 μm. B, Time-lapse microscopy of cells treated with 1 μmol/L AOH1996 or DMSO control. HFK, HeLa, and HeLa POLη cells undergo timely mitotic progression in control conditions. AOH1996 treatment causes extended mitotic arrest and death, particularly in transformed cells. Scale bars = 10 μm. C, Relative cell growth over a 72-hour imaging window. AOH1996 halts proliferation in HFKs and leads to significant cell loss in HeLa and HeLa POLη cells. D, Quantification of time spent in mitosis. AOH1996 significantly prolongs mitosis in all lines, with the greatest delay observed in HeLa POLη cells. E, Time-course of mitotic entry following AOH1996 treatment. All cell types enter mitosis at similar rates. F, Analysis of mitotic outcomes. HFKs primarily undergo single-cell mitotic exit (∼71%), whereas mitotic death is the predominant fate in HeLa and HeLa POLη cells (∼70% to 74%). G, Temporal distribution of single-cell mitotic exits. Most events occur between 20 and 30 hours after treatment in HFKs. H, Temporal profile of mitotic death. HeLa and HeLa POLη cells undergo mitotic death between 30 and 50 hours after treatment. Data represent the mean ± SEM from three independent experiments. Statistical analysis was performed using one-way or two-way ANOVA. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.