Figure 5
From: A preclinical model of cutaneous melanoma based on reconstructed human epidermis
Qualification of mOS-REp as a test system for the assessment of tumor therapeutics. (A) Analysis of metabolic activity. mOS-REpA11 showed no significant difference in viability for different concentrations. In comparison, for OS-REp and mOS-REpSK-MEL-28 weak but statistically relevant decreases of viability were measurable for 5 µM vemurafenib. (B) Influence on proliferation rates. Melanoma cells re-isolated from BRAFWT (mOS-REpA11) and BRAFV600E (mOS-REpSK-MEL-28) melanoma skin equivalents were stained on cytospots (Fig. S2) with the proliferation marker Ki67 and counted quantitatively. In mOS-REpSK-MEL-28 the number of positively stained cells diminished completely after treatment, whereas the Ki67 expression was not altered by vemurafenib treatment in mOS-REpA11. (C) Measurement of glucose consumption. It was either directly measured in the supernatant or visualized locally by fluorescently labelled glucose. Both the fluorescent signal and glucose consumption decreased dose-dependently for mOS-REpSK-MEL-28. This could not be observed in mOS-REpA11. Scale bar 200 µm. (D) Treatment effects on the cell cycle. Cell cycle stages were determined by DNA content measured via flow cytometry after propidium iodide staining. Vemurafenib led to a dose-dependent significant increase of cells in G0 phase and decrease in G2/M phase in mOS-REpSK-MEL-28. OS-REp and mOS-REpA11 showed no treatment-dependent cell cycle response.
