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. 2020 Apr 15;10(2):391–418. doi: 10.1016/j.jcmgh.2020.04.002

Figure 9.

Figure 9

Activation of CSF1R in CRC cells induces migration, invasion, and lung metastases. (A) Genes were preranked by expression correlation coefficient (Pearson r) with CSF1R in descending order from left (positive correlation) to right (negative correlation) based on RNA expression data and association of indicated gene signature with CSF1R expression was subsequently analyzed by GSEA. (B) Scratch assay of DLD1-pRTR-CSF1R cells treated with Dox or Dox/CSF1. Scale bar = 200 μm. (C) Boyden chamber assays of cellular migration (left) or invasion (invasion). (D) SW620 cells were pretreated with inhibitors as indicated, and subsequently treated with Dox and CSF1. After incubation with CSF1 or 48 hours, cells were subjected to a Boyden chamber assay. Cells were transfected with or without pre-miR-34a oligo 1 day before the addition of Dox and CSF1, and then subjected to a (E) migration or (F) invasion assay. (G) DLD1-Luc2/pRTR-CSF1R cells treated with or without Dox and CSF1 were injected into the tail vein of NOD/SCID mice. At the indicated time points, bioluminescence signals were recorded. Bioluminescence signals are presented as “total flux.” (H) Representative examples of bioluminescence imaging at the indicated time points after tail vein injection of DLD1-Luc2/pRTR-CSF1R cells. (I) SW620 cells were transfected with the indicated oligonucleotides for 48 hours and then subjected to an invasion assay in Boyden chambers for another 36 hours. (J) SW620 cells were transfected with the indicated oligonucleotides for 48 hours and subsequently injected into the tail vein of NOD/SCID mice. Left: lungs were resected 8 weeks after injection. Arrows indicate metastatic tumor nodules. Right: representative examples of the hematoxylin and eosin staining of resected lungs are shown. Scale bar = 200 μm. (K) Quantification of metastatic tumor nodules in the lung per mouse 8 weeks after tail-vein injection. In panels B, C, D, E, F, G, I, and K, mean values ± SD are provided. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001.