Epithelial-to-mesenchymal transition in FHC-silenced cells: the role of CXCR4/CXCL12 axis
Abstract Background Ferritin plays a central role in the intracellular iron metabolism; the molecule is a nanocage of 24 subunits of the heavy and light types. The heavy subunit (FHC) is provided of a ferroxidase activity and thus performs the key transformation of iron in a non-toxic form. Recently...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2017-08-01
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| Series: | Journal of Experimental & Clinical Cancer Research |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13046-017-0571-8 |
| _version_ | 1818540398140719104 |
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| author | I. Aversa F. Zolea C. Ieranò S. Bulotta A. M. Trotta M. C. Faniello C. De Marco D. Malanga F. Biamonte G. Viglietto G. Cuda S. Scala F. Costanzo |
| author_facet | I. Aversa F. Zolea C. Ieranò S. Bulotta A. M. Trotta M. C. Faniello C. De Marco D. Malanga F. Biamonte G. Viglietto G. Cuda S. Scala F. Costanzo |
| author_sort | I. Aversa |
| collection | DOAJ |
| description | Abstract Background Ferritin plays a central role in the intracellular iron metabolism; the molecule is a nanocage of 24 subunits of the heavy and light types. The heavy subunit (FHC) is provided of a ferroxidase activity and thus performs the key transformation of iron in a non-toxic form. Recently, it has been shown that FHC is also involved in additional not iron-related critical pathways including, among the others, p53 regulation, modulation of oncomiRNAs expression and chemokine signalling. Epithelial to mesenchymal transition (EMT) is a cellular mechanism by which the cell acquires a fibroblast-like phenotype along with a decreased adhesion and augmented motility. In this work we have focused our attention on the role of the FHC on EMT induction in the human cell lines MCF-7 and H460 to elucidate the underlying molecular mechanisms. Methods Targeted silencing of the FHC was performed by lentiviral-driven shRNA strategy. Reconstitution of the FHC gene product was obtained by full length FHC cDNA transfection with Lipofectamine 2000. MTT and cell count assays were used to evaluate cell viability and proliferation; cell migration capability was assayed by the wound-healing assay and transwell strategy. Quantification of the CXCR4 surface expression was performed by flow cytometry. Results Experimental data indicated that FHC-silenced MCF-7 and H460 cells (MCF-7shFHC, H460shFHC) acquire a mesenchymal phenotype, accompanied by a significant enhancement of their migratory and proliferative capacity. This shift is coupled to an increase in ROS production and by an activation of the CXCR4/CXCL12 signalling pathway. We present experimental data indicating that the cytosolic increase in ROS levels is responsible for the enhanced proliferation of FHC-silenced cells, while the higher migration rate is attributable to a dysregulation of the CXCR4/CXCL12 axis. Conclusions Our findings indicate that induction of EMT, increased migration and survival depend, in MCF-7 and H460 cells, on the release of FHC control on two pathways, namely the iron/ROS metabolism and CXCR4/CXCL12 axis. Besides constituting a further confirmation of the multifunctional nature of FHC, this data also suggest that the analysis of FHC amount/function might be an important additional tool to predict tumor aggressiveness. |
| first_indexed | 2024-12-11T21:54:49Z |
| format | Article |
| id | doaj.art-7c7b11f0cbed470c84dc880814f944dd |
| institution | Directory Open Access Journal |
| issn | 1756-9966 |
| language | English |
| last_indexed | 2024-12-11T21:54:49Z |
| publishDate | 2017-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Experimental & Clinical Cancer Research |
| spelling | doaj.art-7c7b11f0cbed470c84dc880814f944dd2022-12-22T00:49:20ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662017-08-0136111510.1186/s13046-017-0571-8Epithelial-to-mesenchymal transition in FHC-silenced cells: the role of CXCR4/CXCL12 axisI. Aversa0F. Zolea1C. Ieranò2S. Bulotta3A. M. Trotta4M. C. Faniello5C. De Marco6D. Malanga7F. Biamonte8G. Viglietto9G. Cuda10S. Scala11F. Costanzo12Research Center of Advanced Biochemistry and Molecular Biology, Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta CampusResearch Center of Advanced Biochemistry and Molecular Biology, Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta CampusGenomica Funzionale, INT PascaleDepartment of Health Sciences, Magna Græcia University of Catanzaro, Salvatore Venuta CampusGenomica Funzionale, INT PascaleResearch Center of Advanced Biochemistry and Molecular Biology, Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta CampusDepartment of Experimental and Clinical Medicine University of Catanzaro “Magna Graecia”Department of Experimental and Clinical Medicine University of Catanzaro “Magna Graecia”Research Center of Advanced Biochemistry and Molecular Biology, Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta CampusDepartment of Experimental and Clinical Medicine University of Catanzaro “Magna Graecia”Research Center of Advanced Biochemistry and Molecular Biology, Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta CampusGenomica Funzionale, INT PascaleResearch Center of Advanced Biochemistry and Molecular Biology, Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta CampusAbstract Background Ferritin plays a central role in the intracellular iron metabolism; the molecule is a nanocage of 24 subunits of the heavy and light types. The heavy subunit (FHC) is provided of a ferroxidase activity and thus performs the key transformation of iron in a non-toxic form. Recently, it has been shown that FHC is also involved in additional not iron-related critical pathways including, among the others, p53 regulation, modulation of oncomiRNAs expression and chemokine signalling. Epithelial to mesenchymal transition (EMT) is a cellular mechanism by which the cell acquires a fibroblast-like phenotype along with a decreased adhesion and augmented motility. In this work we have focused our attention on the role of the FHC on EMT induction in the human cell lines MCF-7 and H460 to elucidate the underlying molecular mechanisms. Methods Targeted silencing of the FHC was performed by lentiviral-driven shRNA strategy. Reconstitution of the FHC gene product was obtained by full length FHC cDNA transfection with Lipofectamine 2000. MTT and cell count assays were used to evaluate cell viability and proliferation; cell migration capability was assayed by the wound-healing assay and transwell strategy. Quantification of the CXCR4 surface expression was performed by flow cytometry. Results Experimental data indicated that FHC-silenced MCF-7 and H460 cells (MCF-7shFHC, H460shFHC) acquire a mesenchymal phenotype, accompanied by a significant enhancement of their migratory and proliferative capacity. This shift is coupled to an increase in ROS production and by an activation of the CXCR4/CXCL12 signalling pathway. We present experimental data indicating that the cytosolic increase in ROS levels is responsible for the enhanced proliferation of FHC-silenced cells, while the higher migration rate is attributable to a dysregulation of the CXCR4/CXCL12 axis. Conclusions Our findings indicate that induction of EMT, increased migration and survival depend, in MCF-7 and H460 cells, on the release of FHC control on two pathways, namely the iron/ROS metabolism and CXCR4/CXCL12 axis. Besides constituting a further confirmation of the multifunctional nature of FHC, this data also suggest that the analysis of FHC amount/function might be an important additional tool to predict tumor aggressiveness.http://link.springer.com/article/10.1186/s13046-017-0571-8Ferritin heavy chainEMTCXCR4CXCL12CancerROS |
| spellingShingle | I. Aversa F. Zolea C. Ieranò S. Bulotta A. M. Trotta M. C. Faniello C. De Marco D. Malanga F. Biamonte G. Viglietto G. Cuda S. Scala F. Costanzo Epithelial-to-mesenchymal transition in FHC-silenced cells: the role of CXCR4/CXCL12 axis Journal of Experimental & Clinical Cancer Research Ferritin heavy chain EMT CXCR4 CXCL12 Cancer ROS |
| title | Epithelial-to-mesenchymal transition in FHC-silenced cells: the role of CXCR4/CXCL12 axis |
| title_full | Epithelial-to-mesenchymal transition in FHC-silenced cells: the role of CXCR4/CXCL12 axis |
| title_fullStr | Epithelial-to-mesenchymal transition in FHC-silenced cells: the role of CXCR4/CXCL12 axis |
| title_full_unstemmed | Epithelial-to-mesenchymal transition in FHC-silenced cells: the role of CXCR4/CXCL12 axis |
| title_short | Epithelial-to-mesenchymal transition in FHC-silenced cells: the role of CXCR4/CXCL12 axis |
| title_sort | epithelial to mesenchymal transition in fhc silenced cells the role of cxcr4 cxcl12 axis |
| topic | Ferritin heavy chain EMT CXCR4 CXCL12 Cancer ROS |
| url | http://link.springer.com/article/10.1186/s13046-017-0571-8 |
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