Fig. 5
Cells and organoids from drug-resistant CRC patients rely on the WNT/β‐catenin pathway. a Primary 2D cell lines, established from the tissue specimen collected from a CRC patient whose tumour developed secondary resistance to anti-EGFR therapy (see detailed methods section), were electroporated with a plasmid encoding for WT APC or an inactive APC version (G97*). Electroporation buffer was used as the control (mock). After 48 h, cells were stained with Hoechst 3342 /Propidium Iodide (PI) to detect cell death. A representative image of a single 96-well is shown for each condition. b Relative quantification of Hoechst/PI positive cells was made using ImageJ software and normalised against mock cells. Results represent means ± SD of three independent wells. c 3D organoids established from a CRC patient whose tumour developed secondary resistance to EGFR-BRAF combinatorial treatment (see detailed methods section), were treated with LGK974 for 2 weeks. Representative confocal microscopy images showing active cleaved caspase-3 (green) are shown. Nuclei are stained with DAPI (blue) and actin with Phalloidin (red). Maximum projection of a 10 image stack along the z-axis. Scale bar: 50 μm. d Patient-derived mice models (xenopatient) were established from a tumour obtained from a metastatic colorectal cancer patient (PZ-2) resistant to EGFR/BRAF combinatorial treatment. Upon successful engraftment, mice were randomised to vehicle (n = 6) or LGK974 (WNT inhibitor) (n = 6) treated arm. Results represent tumour mass volume (mm3, mean ± CI of individual tumour volume)
