Figure 4
From: Distinct routes to metastasis: plasticity-dependent and plasticity-independent pathways
AT3 and DU145 cells metastasize via MET-independent pathways. (a) DTs and AT3 cells were left untransfected or transfected with an empty vector (pcDNA6) or E-cadDipIIIcI2 and subsequently selected with blasticidin. All untransfected DT and AT3 cells are killed by blasticidin, while DTs and AT3 cells transfected with an empty vector containing a blasticidin resistance marker survive blasticidin selection. Importantly, only mesenchymal AT3 cells transfected with the E-cadDipIIIcI2 suicide reporter grow out during blasticidin selection, while all epithelial DT cells with the reporter die. (b) Quantification of DT and AT3 growth during blasticidin selection by WST-1 cell growth assay. (c) Representative lung metastases from AT3 cells harboring RIIIcI2+Gint or E-cadDipIIIcI2+Gint. (d) No difference was observed in the number of macrometastases in each group. (e) Injection of DU145 E-cadCreIIIcI2 (as a control) or E-cadDipIIIcI2 cells led to formation of lung metastases with 100% penetrance. There was no difference in the number of metastases between control cells or cells with the suicide reporter. (f) DU145 E-cadDipIIIcI2 metastases lack E-cadherin expression and stain positive for vimentin.
