Actin cytoskeleton regulation of epithelial mesenchymal transition in metastatic cancer cells.
Epithelial-mesenchymal transition (EMT) is associated with loss of the cell-cell adhesion molecule E-cadherin and disruption of cell-cell junctions as well as with acquisition of migratory properties including reorganization of the actin cytoskeleton and activation of the RhoA GTPase. Here we show t...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4355409?pdf=render |
| _version_ | 1818229760050855936 |
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| author | Jay Shankar Ivan R Nabi |
| author_facet | Jay Shankar Ivan R Nabi |
| author_sort | Jay Shankar |
| collection | DOAJ |
| description | Epithelial-mesenchymal transition (EMT) is associated with loss of the cell-cell adhesion molecule E-cadherin and disruption of cell-cell junctions as well as with acquisition of migratory properties including reorganization of the actin cytoskeleton and activation of the RhoA GTPase. Here we show that depolymerization of the actin cytoskeleton of various metastatic cancer cell lines with Cytochalasin D (Cyt D) reduces cell size and F-actin levels and induces E-cadherin expression at both the protein and mRNA level. Induction of E-cadherin was dose dependent and paralleled loss of the mesenchymal markers N-cadherin and vimentin. E-cadherin levels increased 2 hours after addition of Cyt D in cells showing an E-cadherin mRNA response but only after 10-12 hours in HT-1080 fibrosarcoma and MDA-MB-231 cells in which E-cadherin mRNA level were only minimally affected by Cyt D. Cyt D treatment induced the nuclear-cytoplasmic translocation of EMT-associated SNAI 1 and SMAD1/2/3 transcription factors. In non-metastatic MCF-7 breast cancer cells, that express E-cadherin and represent a cancer cell model for EMT, actin depolymerization with Cyt D induced elevated E-cadherin while actin stabilization with Jasplakinolide reduced E-cadherin levels. Elevated E-cadherin levels due to Cyt D were associated with reduced activation of Rho A. Expression of dominant-negative Rho A mutant increased and dominant-active Rho A mutant decreased E-cadherin levels and also prevented Cyt D induction of E-cadherin. Reduced Rho A activation downstream of actin remodelling therefore induces E-cadherin and reverses EMT in cancer cells. Cyt D treatment inhibited migration and, at higher concentrations, induced cytotoxicity of both HT-1080 fibrosarcoma cells and normal Hs27 fibroblasts, but only induced mesenchymal-epithelial transition in HT-1080 cancer cells. Our studies suggest that actin remodelling is an upstream regulator of EMT in metastatic cancer cells. |
| first_indexed | 2024-12-12T10:23:43Z |
| format | Article |
| id | doaj.art-10976116972049ab992ab62957afe3e5 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-12T10:23:43Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-10976116972049ab992ab62957afe3e52022-12-22T00:27:29ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01103e011995410.1371/journal.pone.0119954Actin cytoskeleton regulation of epithelial mesenchymal transition in metastatic cancer cells.Jay ShankarIvan R NabiEpithelial-mesenchymal transition (EMT) is associated with loss of the cell-cell adhesion molecule E-cadherin and disruption of cell-cell junctions as well as with acquisition of migratory properties including reorganization of the actin cytoskeleton and activation of the RhoA GTPase. Here we show that depolymerization of the actin cytoskeleton of various metastatic cancer cell lines with Cytochalasin D (Cyt D) reduces cell size and F-actin levels and induces E-cadherin expression at both the protein and mRNA level. Induction of E-cadherin was dose dependent and paralleled loss of the mesenchymal markers N-cadherin and vimentin. E-cadherin levels increased 2 hours after addition of Cyt D in cells showing an E-cadherin mRNA response but only after 10-12 hours in HT-1080 fibrosarcoma and MDA-MB-231 cells in which E-cadherin mRNA level were only minimally affected by Cyt D. Cyt D treatment induced the nuclear-cytoplasmic translocation of EMT-associated SNAI 1 and SMAD1/2/3 transcription factors. In non-metastatic MCF-7 breast cancer cells, that express E-cadherin and represent a cancer cell model for EMT, actin depolymerization with Cyt D induced elevated E-cadherin while actin stabilization with Jasplakinolide reduced E-cadherin levels. Elevated E-cadherin levels due to Cyt D were associated with reduced activation of Rho A. Expression of dominant-negative Rho A mutant increased and dominant-active Rho A mutant decreased E-cadherin levels and also prevented Cyt D induction of E-cadherin. Reduced Rho A activation downstream of actin remodelling therefore induces E-cadherin and reverses EMT in cancer cells. Cyt D treatment inhibited migration and, at higher concentrations, induced cytotoxicity of both HT-1080 fibrosarcoma cells and normal Hs27 fibroblasts, but only induced mesenchymal-epithelial transition in HT-1080 cancer cells. Our studies suggest that actin remodelling is an upstream regulator of EMT in metastatic cancer cells.http://europepmc.org/articles/PMC4355409?pdf=render |
| spellingShingle | Jay Shankar Ivan R Nabi Actin cytoskeleton regulation of epithelial mesenchymal transition in metastatic cancer cells. PLoS ONE |
| title | Actin cytoskeleton regulation of epithelial mesenchymal transition in metastatic cancer cells. |
| title_full | Actin cytoskeleton regulation of epithelial mesenchymal transition in metastatic cancer cells. |
| title_fullStr | Actin cytoskeleton regulation of epithelial mesenchymal transition in metastatic cancer cells. |
| title_full_unstemmed | Actin cytoskeleton regulation of epithelial mesenchymal transition in metastatic cancer cells. |
| title_short | Actin cytoskeleton regulation of epithelial mesenchymal transition in metastatic cancer cells. |
| title_sort | actin cytoskeleton regulation of epithelial mesenchymal transition in metastatic cancer cells |
| url | http://europepmc.org/articles/PMC4355409?pdf=render |
| work_keys_str_mv | AT jayshankar actincytoskeletonregulationofepithelialmesenchymaltransitioninmetastaticcancercells AT ivanrnabi actincytoskeletonregulationofepithelialmesenchymaltransitioninmetastaticcancercells |