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Mesenchymal-transitioned cancer cells instigate the invasion of epithelial cancer cells through secretion of WNT3 and WNT5B.

Kato S, Hayakawa Y, Sakurai H, Saiki I, Yokoyama S - Cancer Sci. (2014)

Bottom Line: In this study, we show that the mesenchymal-transitioned cancer cells instigate the invasive ability and metastatic potential of the neighboring epithelial cancer cells in vitro and in vivo.We identify WNT3 and WNT5B as critical factors secreted from Transforming growth factor-induced mesenchymal cancer cells for instigating the epithelial cancer cell invasion along with the induction of secondary EMT phenotype.These results shed light on the significance of cancer heterogeneity and the interaction between epithelial and mesenchymal-transitioned cancer cells within the tumor microenvironment in promoting metastatic disease through the WNT-dependent mechanism.

View Article: PubMed Central - PubMed

Affiliation: Division of Pathogenic Biochemistry, Institute of Natural Medicine, University of Toyama, Toyama, Japan.

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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.
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fig02: 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.

Mentions: To determine whether the direct cell–cell interaction is required for promoting the metastatic potential of E-cells by neighboring with M-cells, we first examined the expression of EMT-related proteins in A549 or Panc-1 cells under the co-culture of E-cells with M-cells in the presence or absence of direct cell–cell contact (Fig.2a,b). Given that the upregulation of the mesenchymal marker (Snail and Vimentin) and the downregulation of the epithelial marker (E-cadherin) were markedly induced by both direct and separated co-culture of E-cells and M-cells, the soluble factor(s) derived from M-cells was at least sufficient for the induction of the secondary EMT phenotype of E-cells upon co-culture with M-cells. We further confirmed that the invasion of E-A549 cells was remarkably enhanced after the culture with M-A549-derived conditioned medium (M-A549-CM) compared with E-A549-CM (Fig.2c and Fig. S3). In concert with the enhanced invasive ability of E-A549 cells after the cultivation with M-A549-CM, the secondary EMT phenotype in E-A549 cells was also induced after the culture with M-A549-CM. Similar results were obtained in the invasion of E-Panc cells after the cultivation with M-Panc-CM (data not shown).


Mesenchymal-transitioned cancer cells instigate the invasion of epithelial cancer cells through secretion of WNT3 and WNT5B.

Kato S, Hayakawa Y, Sakurai H, Saiki I, Yokoyama S - Cancer Sci. (2014)

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.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317934&req=5

fig02: 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.
Mentions: To determine whether the direct cell–cell interaction is required for promoting the metastatic potential of E-cells by neighboring with M-cells, we first examined the expression of EMT-related proteins in A549 or Panc-1 cells under the co-culture of E-cells with M-cells in the presence or absence of direct cell–cell contact (Fig.2a,b). Given that the upregulation of the mesenchymal marker (Snail and Vimentin) and the downregulation of the epithelial marker (E-cadherin) were markedly induced by both direct and separated co-culture of E-cells and M-cells, the soluble factor(s) derived from M-cells was at least sufficient for the induction of the secondary EMT phenotype of E-cells upon co-culture with M-cells. We further confirmed that the invasion of E-A549 cells was remarkably enhanced after the culture with M-A549-derived conditioned medium (M-A549-CM) compared with E-A549-CM (Fig.2c and Fig. S3). In concert with the enhanced invasive ability of E-A549 cells after the cultivation with M-A549-CM, the secondary EMT phenotype in E-A549 cells was also induced after the culture with M-A549-CM. Similar results were obtained in the invasion of E-Panc cells after the cultivation with M-Panc-CM (data not shown).

Bottom Line: In this study, we show that the mesenchymal-transitioned cancer cells instigate the invasive ability and metastatic potential of the neighboring epithelial cancer cells in vitro and in vivo.We identify WNT3 and WNT5B as critical factors secreted from Transforming growth factor-induced mesenchymal cancer cells for instigating the epithelial cancer cell invasion along with the induction of secondary EMT phenotype.These results shed light on the significance of cancer heterogeneity and the interaction between epithelial and mesenchymal-transitioned cancer cells within the tumor microenvironment in promoting metastatic disease through the WNT-dependent mechanism.

View Article: PubMed Central - PubMed

Affiliation: Division of Pathogenic Biochemistry, Institute of Natural Medicine, University of Toyama, Toyama, Japan.

Show MeSH
Related in: MedlinePlus