Overexpression and Tyr421-phosphorylation of cortactin is induced by three-dimensional spheroid culturing and contributes to migration and invasion of pancreatic ductal adenocarcinoma (PDAC) cells
Abstract Background The nucleation-promoting factor cortactin is expressed and promotes tumor progression and metastasis in various cancers. However, little is known about the biological role of cortactin in the progression of pancreatic ductal adenocarcinoma (PDAC). Methods Cortactin and phosphoryl...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-03-01
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| Series: | Cancer Cell International |
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| Online Access: | http://link.springer.com/article/10.1186/s12935-019-0798-x |
| _version_ | 1818216407109730304 |
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| author | Katharina Stock Rebekka Borrink Jan-Henrik Mikesch Anna Hansmeier Jan Rehkämper Marcel Trautmann Eva Wardelmann Wolfgang Hartmann Jan Sperveslage Konrad Steinestel |
| author_facet | Katharina Stock Rebekka Borrink Jan-Henrik Mikesch Anna Hansmeier Jan Rehkämper Marcel Trautmann Eva Wardelmann Wolfgang Hartmann Jan Sperveslage Konrad Steinestel |
| author_sort | Katharina Stock |
| collection | DOAJ |
| description | Abstract Background The nucleation-promoting factor cortactin is expressed and promotes tumor progression and metastasis in various cancers. However, little is known about the biological role of cortactin in the progression of pancreatic ductal adenocarcinoma (PDAC). Methods Cortactin and phosphorylated cortactin (Y421) were investigated immunohistochemically in 66 PDAC tumor specimens. To examine the functional role of cortactin in PDAC, we modulated cortactin expression by establishing two cortactin knockout cell lines (Panc-1 and BxPC-3) with CRISPR/Cas9 technique. Cortactin knockout was verified by immunoblotting and immunofluorescence microscopy and functional effects were determined by cell migration and invasion assays. A proteomic screening approach was performed to elucidate potential binding partners of cortactin. Results Immunohistochemically, we observed higher cortactin expression and Tyr421-phosphorylation in PDAC metastases compared to primary tumor tissues. In PDAC cell lines Panc-1 and BxPC-3, knockdown of cortactin impaired migration and invasion, while cell proliferation was not affected. Three-dimensional spheroid culturing as a model for collective cell migration enhanced cortactin expression and Tyr421-phosphorylation. The activation of cortactin as well as the migratory capacity of PDAC cells could significantly be reduced by dasatinib, a Src family kinase inhibitor. Finally, we identified gelsolin as a novel protein interaction partner of cortactin in PDAC. Conclusion Our data provides evidence that cohesive cell migration induces cortactin expression and phosphorylation as a prerequisite for the gain of an invasive, pro-migratory phenotype in PDAC that can effectively be targeted with dasatinib. |
| first_indexed | 2024-12-12T06:51:29Z |
| format | Article |
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| language | English |
| last_indexed | 2024-12-12T06:51:29Z |
| publishDate | 2019-03-01 |
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| series | Cancer Cell International |
| spelling | doaj.art-b8529c2a6daf4252982af6a08f97ac172022-12-22T00:34:03ZengBMCCancer Cell International1475-28672019-03-0119111510.1186/s12935-019-0798-xOverexpression and Tyr421-phosphorylation of cortactin is induced by three-dimensional spheroid culturing and contributes to migration and invasion of pancreatic ductal adenocarcinoma (PDAC) cellsKatharina Stock0Rebekka Borrink1Jan-Henrik Mikesch2Anna Hansmeier3Jan Rehkämper4Marcel Trautmann5Eva Wardelmann6Wolfgang Hartmann7Jan Sperveslage8Konrad Steinestel9Gerhard-Domagk-Institute of Pathology, University Hospital MünsterGerhard-Domagk-Institute of Pathology, University Hospital MünsterDepartment of Medicine A, University Hospital MünsterDepartment of Medicine A, University Hospital MünsterGerhard-Domagk-Institute of Pathology, University Hospital MünsterGerhard-Domagk-Institute of Pathology, University Hospital MünsterGerhard-Domagk-Institute of Pathology, University Hospital MünsterGerhard-Domagk-Institute of Pathology, University Hospital MünsterGerhard-Domagk-Institute of Pathology, University Hospital MünsterGerhard-Domagk-Institute of Pathology, University Hospital MünsterAbstract Background The nucleation-promoting factor cortactin is expressed and promotes tumor progression and metastasis in various cancers. However, little is known about the biological role of cortactin in the progression of pancreatic ductal adenocarcinoma (PDAC). Methods Cortactin and phosphorylated cortactin (Y421) were investigated immunohistochemically in 66 PDAC tumor specimens. To examine the functional role of cortactin in PDAC, we modulated cortactin expression by establishing two cortactin knockout cell lines (Panc-1 and BxPC-3) with CRISPR/Cas9 technique. Cortactin knockout was verified by immunoblotting and immunofluorescence microscopy and functional effects were determined by cell migration and invasion assays. A proteomic screening approach was performed to elucidate potential binding partners of cortactin. Results Immunohistochemically, we observed higher cortactin expression and Tyr421-phosphorylation in PDAC metastases compared to primary tumor tissues. In PDAC cell lines Panc-1 and BxPC-3, knockdown of cortactin impaired migration and invasion, while cell proliferation was not affected. Three-dimensional spheroid culturing as a model for collective cell migration enhanced cortactin expression and Tyr421-phosphorylation. The activation of cortactin as well as the migratory capacity of PDAC cells could significantly be reduced by dasatinib, a Src family kinase inhibitor. Finally, we identified gelsolin as a novel protein interaction partner of cortactin in PDAC. Conclusion Our data provides evidence that cohesive cell migration induces cortactin expression and phosphorylation as a prerequisite for the gain of an invasive, pro-migratory phenotype in PDAC that can effectively be targeted with dasatinib.http://link.springer.com/article/10.1186/s12935-019-0798-xPancreatic ductal adenocarcinomaCortactinTumor cell migration and invasion |
| spellingShingle | Katharina Stock Rebekka Borrink Jan-Henrik Mikesch Anna Hansmeier Jan Rehkämper Marcel Trautmann Eva Wardelmann Wolfgang Hartmann Jan Sperveslage Konrad Steinestel Overexpression and Tyr421-phosphorylation of cortactin is induced by three-dimensional spheroid culturing and contributes to migration and invasion of pancreatic ductal adenocarcinoma (PDAC) cells Cancer Cell International Pancreatic ductal adenocarcinoma Cortactin Tumor cell migration and invasion |
| title | Overexpression and Tyr421-phosphorylation of cortactin is induced by three-dimensional spheroid culturing and contributes to migration and invasion of pancreatic ductal adenocarcinoma (PDAC) cells |
| title_full | Overexpression and Tyr421-phosphorylation of cortactin is induced by three-dimensional spheroid culturing and contributes to migration and invasion of pancreatic ductal adenocarcinoma (PDAC) cells |
| title_fullStr | Overexpression and Tyr421-phosphorylation of cortactin is induced by three-dimensional spheroid culturing and contributes to migration and invasion of pancreatic ductal adenocarcinoma (PDAC) cells |
| title_full_unstemmed | Overexpression and Tyr421-phosphorylation of cortactin is induced by three-dimensional spheroid culturing and contributes to migration and invasion of pancreatic ductal adenocarcinoma (PDAC) cells |
| title_short | Overexpression and Tyr421-phosphorylation of cortactin is induced by three-dimensional spheroid culturing and contributes to migration and invasion of pancreatic ductal adenocarcinoma (PDAC) cells |
| title_sort | overexpression and tyr421 phosphorylation of cortactin is induced by three dimensional spheroid culturing and contributes to migration and invasion of pancreatic ductal adenocarcinoma pdac cells |
| topic | Pancreatic ductal adenocarcinoma Cortactin Tumor cell migration and invasion |
| url | http://link.springer.com/article/10.1186/s12935-019-0798-x |
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