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. 2017 Nov 11;8(64):107907–107919. doi: 10.18632/oncotarget.22392

Figure 4. Small CRC cells possess higher metastatic capacity.

Figure 4

(A-B) Transwell assays. Large- and small-sized subpopulations were sorted out in LoVo, HT-29 and xhCRC cells, and seeded in transwell inserts covered with Matrigel. After 24 hours, cells migrated through Matrigel barrier were photographed (left panel) and calculated (right panel). Scale bars: 100 μm. Data are presented from triple experiments. (C-D) Sorted large and small LoVo cells were injected into the tail vein of NOD/SCID mice (n=6 per group). 8 weeks later, animals were scarified for examining metastatic lesions in the lungs. Expression of CK20 and EpCAM in metastatic lesions was examined by immunohistochemistry staining. Scale bars: 100 μm. (E-F) Purified large and small LoVo cells were implanted subcutaneously into BALB/c-nu female. 6 weeks later, the mice were scarified for harvesting the lungs to examine metastatic lesions. Implanted at 10K cell dose, both large and small LoVo cells formed metastatic lesions in the lungs of the mice, while at 1K cell dose, only small LoVo cells formed lung metastatic lesions. Expression of CK20 and EpCAM in metastatic lesions was confirmed by immunohistochemistry staining. Scale bars: 100 μm.(Mean ±SD, n =5, *P< 0.05, **P< 0.01, ***P< 0.001)