An Ovol2-Zeb1 Mutual Inhibitory Circuit Governs Bidirectional and Multi-step Transition between Epithelial and Mesenchymal States
Fig 3
Experimental evidence for bidirectional potential of MCF10A cells.
A, B, D, E) Flow cytometric analysis of epithelial marker (Ecad) and mesenchymal marker (Vim) profiles. A) Direct comparison of MCF10A with luminal (epithelial)-type cancer cell line MCF7 and basal (mesenchymal)-type cancer cell line MDA-MB231. MCF10A (green) falls in the intermediate state between MCF7 (blue) and MDA-MB231 (red). Analyses were performed at 90–100% confluency. B) Bidirectional potential of MCF10A cells. E(I)MT and M(I)ET was induced by forced expression of transcription factors Snail or Zeb1, and Ovol2, respectively. After 6 days of lentiviral infection, Snail (red) and Zeb1 (orange) induced EMT while Ovol2 (blue) induced MET as compared to the empty vector control (green). C) Stochastic simulations for a population of 2000 cells in three different conditions: basal parameter set (green), high basal production rate of Zeb1 ZEB1 (red, Zeb1 mRNA basal production rate was raised to 0.01 μM/hr) and high basal production rate of Ovol2 (blue, Ovol2 basal production rate was raised to 2 μM/hr). Initial conditions are all at I1 state. D) The histogram shows the expression status of CD44. “M” states (Snail;red and Zeb1;orange) correlate with high CD44 expression while cells in “E” state (Ovol2;blue) show low CD44 expression as compared to empty vector control (green). MCF7 is shown as a representative cell type in the “E” state with low CD44 expression. E) Ovol2 reprograms MDA-MB231 cells from M- to E- state. Cells were analyzed after 6 days of control (red) or Ovol2-expressing (blue) lentiviral infection. F) Stochastic simulations with a basal parameter set (red) and high basal production rate of Ovol2 (blue, Ovol2 basal production rate was raised to 2 μM/hr). Initial conditions are all at M state.