Journal: Cancer Science

Article Title: Mesenchymal-transitioned cancer cells instigate the invasion of epithelial cancer cells through secretion of WNT3 and WNT5B

doi: 10.1111/cas.12336

Figure Lengend Snippet: Cell–cell contact independent induction of secondary epithelial-to-mesenchymal transition (EMT) phenotype and invasiveness in neighboring epithelial cancer cells by mesenchymal-transitioned cancer cells. (a, b) A549 cells were co-cultured directly (a) or separately in transwell cell culture chamber (b) at the indicated cell numbers for 24 h and EMT-related protein expression were determined by western blotting. No cells (−), E-cells (E) or M-cells (M) were seeded in upper compartment of transwell chamber. In the separated co-culture, the total protein from E-cells in lower compartment was examined. (c, d) Epithelial A549 cells were treated with conditioned mediums (CM) from E-/M-A549 cells (c) or E-/M-Panc cells (d) for 48 h and subjected to Matrigel invasion assay or western blotting. Total invaded cells were counted after H&E staining. Data represented as the mean ± SD of triplicate experiment. ** P < 0.01 versus E-CM group by two-tailed Student's t test.

Article Snippet: For the direct co-culture experiment, E-cells and M-cells were re-seeded into 60 mm culture dish according to the indicated cell number and co-cultured for 24 h. For the separated co-culture experiment, 1 × 10 5 E-cells and 3 × 10 5 cells of M-cells were seeded into lower or upper compartments of the transwell chamber with 1 μm pore diameter (BD Falcon, Bedford, MA, USA) for 24 h.

Techniques: Cell Culture, Expressing, Western Blot, Co-Culture Assay, Invasion Assay, Staining, Two Tailed Test