Fig. 1.

Statins inhibit cSCC cell growth in a dose-dependent manner through apoptosis and reduce 3D spatial organization. A431 cells (5000 cells/well) were incubated with increasing doses of (A) atorvastatin and (B) simvastatin. Cells incubated with statins showed decreased viability in a dose-dependent manner when measured with ELISA. Each experiment was performed in triplicate and repeated at least 3 times. Statistical analyses were performed with repeated measures ANOVA, which showed significance for atorvastatin (p = 0.00050) and simvastatin (p = 0.00000000001293). IC50 estimated values were 90µM for atorvastatin and 30µM for simvastatin. Annexin V FACS showed a dose-dependent increase in apoptosis for cSCC (C) cells exposed to statins (25,000 cells/well). When compared with control, there was (D) a significant increase in overall apoptosis for 20µM simvastatin (p = 0.0046), 40µM simvastatin (p = 0.0005), 80µM atorvastatin (p = 0.007), and 160µM atorvastatin (p = 0.001). All viable droplets were measured (n = 1,283). All treatment groups showed (E) a significant decrease in spheroid formation when compared with control. Statistical analyses were performed with a two-tailed t-test. **p < 0.01, ***p < 0.001 and ****p < 0.0001.