FIGURE 8:

OCMetSim-Single Cells model with added cell death from crowding predicts that cell crowding and cell death from crowding affect the number and rate at which ovarian cancer cells reach a distant niche site. (A, B) Single-cell simulations run to investigate the effects of cells dying from nonattachment caused by crowding of cells lining the blood vessel. The probability of cell death from crowding was varied and tested. Eighty percent probability of cell death in the blood was used. Rac1 CRISPR-Cas9 KD resulted in a further decrease in the number of cells that reached a distant metastatic niche. (B) The rate at which the Rac1 OE cells reached the distant niche site was decreased with crowding or crowding cell death, and the Rac1 OE reached the distant site faster than the Rac1 CRISPR-Cas9 KD cells. (C,D) Single-cell simulations with 5000 cells and cell crowding were run. (C) Crowding did not affect the number of cells that reached the distant tissue site. However, with 50% probability of dying due to cell crowding, the numbers of Rac1 OE and KD cells that reached the distant site were decreased. (D) The rate at which the Rac1 OE and Rac1 CRISPR-Cas9 KD clone 26 cells entered the distant niche site was also decreased with crowding and crowding cell death. Thirty simulations were run for each experiment. Error bars represent means ± SD. Two-way ANOVA followed by Tukey’s post-hoc test for multiple comparison across columns and rows was used to compare cell types and crowding and crowding death: *p < 0.05,***p < 0.005, ****p < 0.0001.