Fig. 1: Epithelial, hybrid E/M, and mesenchymal cancer cells are detected in mouse and patient cSCCs.
From: Cancer cell plasticity defines response to immunotherapy in cutaneous squamous cell carcinoma
a Representative flow cytometry profile of α6-integrin+CD45−EpCAM+ and EpCAM− cancer cells within WD-SCCs, MD/PD-SCCs, and PD/S-SCCs, which had previously been generated by orthotopic serial engraftments8. b Percentage of mesenchymal GFP+CD45−EpCAM− cancer cells generated after engrafting the indicated GFP+ cancer cells into immunocompetent syngeneic mice (n = 52 tumors per group). c Flow cytometry strategy to isolate full epithelial, EpCAMhigh, EpCAMlow, EpCAM− and full mesenchymal cancer cells from WD-SCCs, MD/PD-SCCs, and PD/S-SCCs based on EpCAM expression. d, e mRNA expression levels of d epithelial differentiation and e EMT genes in the indicated cancer cells relative to full epithelial cancer cells (n = 3 biologically independent samples per group). dNp63, Zeb2 (****p < 0.0001), Krt14 (**p = 0.0010, **p = 0.0011), Grhl1 (*p = 0.0156, ****p < 0.0001, ***p = 0.0002), Grhl2 (***p = 0.0005, ****p < 0.0001), Epcam (**p = 0.0030, ****p < 0.0001), Ovol1 (***p = 0.0001, ****p < 0.0001), Ovol2 (**p = 0.0038, **p = 0.0087), Cdh1 (**p = 0.0034, ****p < 0.0001), Vim (*p = 0.0161, ****p < 0.0001), Snail (***p = 0.0004, ****p < 0.0001), Twist (*p = 0.0333, ***p = 0.0002, ****p < 0.0001), Zeb1 (***p = 0.0002, ****p < 0.0001). f Percentage of EpCAMhigh, EpCAMlow, and EpCAM− cancer cells within tumors generated after engrafting full epithelial (n = 41), EpCAMhigh (n = 32), EpCAMlow (n = 10), and EpCAM− (n = 49) cancer cells. g Representative immunofluorescence images of Ecad+ (red), Vim+ (green), and DAPI nuclear (blue) staining in G2, G3, and G4 patient cSCCs. Scale bar, 100 µm. h–j Quantification of h epithelial Ecad+Vim−, i mesenchymal Ecad−Vim+, and j hybrid Ecad+Vim+ cancer cells per tumor area (mm2) in G2 (n = 4), G3 (n = 6), and G4 (n = 4) patient cSCCs. Each dot represents the average quantification of at least seven fields from different tumor regions. k Percentage of Ecad+Vim−, Ecad+Vim+, and Ecad−Vim+ cancer cells relative to total cancer cells in epithelial (n = 4), mixed (n = 6), and mesenchymal (n = 4) patient cSCCs. Data are represented as the mean ± SD (b, d–f) or ± SEM (h–k), and n values indicate independent tumors (b, f, h–k). P values are determined by one-way ANOVA with Tukey’s (b, h–j) or Dunnett’s (d, e) multiple comparison tests. See Supplementary Fig. 2 for the gating strategy (b, f). Source data are provided as a Source Data file.
