Expression Changes and Impact of the Extracellular Matrix on Etoposide Resistant Human Retinoblastoma Cell Lines
Retinoblastoma (RB) represents the most common malignant childhood eye tumor worldwide. Several studies indicate that the extracellular matrix (ECM) plays a crucial role in tumor growth and metastasis. Moreover, recent studies indicate that the ECM composition might influence the development of resi...
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MDPI AG
2020-06-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/12/4322 |
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| author | Jacqueline Reinhard Natalie Wagner Miriam M. Krämer Marvin Jarocki Stephanie C. Joachim H. Burkhard Dick Andreas Faissner Vinodh Kakkassery |
| author_facet | Jacqueline Reinhard Natalie Wagner Miriam M. Krämer Marvin Jarocki Stephanie C. Joachim H. Burkhard Dick Andreas Faissner Vinodh Kakkassery |
| author_sort | Jacqueline Reinhard |
| collection | DOAJ |
| description | Retinoblastoma (RB) represents the most common malignant childhood eye tumor worldwide. Several studies indicate that the extracellular matrix (ECM) plays a crucial role in tumor growth and metastasis. Moreover, recent studies indicate that the ECM composition might influence the development of resistance to chemotherapy drugs. The objective of this study was to evaluate possible expression differences in the ECM compartment of the parental human cell lines WERI-RB1 (retinoblastoma 1) and Y79 and their Etoposide resistant subclones via polymerase chain reaction (PCR). Western blot analyses were performed to analyze protein levels. To explore the influence of ECM molecules on RB cell proliferation, death, and cluster formation, WERI-RB1 and resistant WERI-ETOR cells were cultivated on Fibronectin, Laminin, Tenascin-C, and Collagen IV and analyzed via time-lapse video microscopy as well as immunocytochemistry. We revealed a significantly reduced mRNA expression of the proteoglycans <i>Brevican</i>, <i>Neurocan,</i> and <i>Versican</i> in resistant WERI-ETOR compared to sensitive WERI-RB1 cells. Also, for the glycoproteins <i>α1-Laminin</i>, <i>Fibronectin</i>, <i>Tenascin-C,</i> and <i>Tenascin-R</i> as well as <i>Collagen IV</i>, reduced expression levels were observed in WERI-ETOR. Furthermore, a downregulation was detected for the matrix metalloproteinases <i>MMP2</i>, <i>MMP7</i>, <i>MMP9</i>, the tissue-inhibitor of metalloproteinase <i>TIMP2</i>, the Integrin receptor subunits <i>ITGA4</i>, <i>ITGA5</i> and <i>ITGB1,</i> and all <i>receptor protein tyrosine phosphatase β/ζ</i> isoforms. Downregulation of <i>Brevican</i>, <i>Collagen IV</i>, <i>Tenascin-R</i>, <i>MMP2</i>, <i>TIMP2,</i> and <i>ITGA5</i> was also verified in Etoposide resistant Y79 cells compared to sensitive ones. Protein levels of Tenascin-C and MMP-2 were comparable in both WERI cell lines. Interestingly, Fibronectin displayed an apoptosis-inducing effect on WERI-RB1 cells, whereas an anti-apoptotic influence was observed for Tenascin-C. Conversely, proliferation of WERI-ETOR cells was enhanced on Tenascin-C, while an anti-proliferative effect was observed on Fibronectin. In WERI-ETOR, cluster formation was decreased on the substrates Collagen IV, Fibronectin, and Tenascin-C. Collectively, we noted a different ECM mRNA expression and behavior of Etoposide resistant compared to sensitive RB cells. These findings may indicate a key role of ECM components in chemotherapy resistance formation of RB. |
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| spelling | doaj.art-c0f83822fe45403d8c1008186c5335772023-11-20T04:10:14ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-06-012112432210.3390/ijms21124322Expression Changes and Impact of the Extracellular Matrix on Etoposide Resistant Human Retinoblastoma Cell LinesJacqueline Reinhard0Natalie Wagner1Miriam M. Krämer2Marvin Jarocki3Stephanie C. Joachim4H. Burkhard Dick5Andreas Faissner6Vinodh Kakkassery7Department of Cell Morphology and Molecular Neurobiology, Faculty of Biology and Biotechnology, Ruhr-University Bochum, Universitaetsstrasse 150, 44780 Bochum, GermanyDepartment of Cell Morphology and Molecular Neurobiology, Faculty of Biology and Biotechnology, Ruhr-University Bochum, Universitaetsstrasse 150, 44780 Bochum, GermanyDepartment of Cell Morphology and Molecular Neurobiology, Faculty of Biology and Biotechnology, Ruhr-University Bochum, Universitaetsstrasse 150, 44780 Bochum, GermanyDepartment of Cell Morphology and Molecular Neurobiology, Faculty of Biology and Biotechnology, Ruhr-University Bochum, Universitaetsstrasse 150, 44780 Bochum, GermanyExperimental Eye Research Institute, University Eye Hospital, Ruhr-University Bochum, In der Schornau 23-25, 44892 Bochum, GermanyExperimental Eye Research Institute, University Eye Hospital, Ruhr-University Bochum, In der Schornau 23-25, 44892 Bochum, GermanyDepartment of Cell Morphology and Molecular Neurobiology, Faculty of Biology and Biotechnology, Ruhr-University Bochum, Universitaetsstrasse 150, 44780 Bochum, GermanyExperimental Eye Research Institute, University Eye Hospital, Ruhr-University Bochum, In der Schornau 23-25, 44892 Bochum, GermanyRetinoblastoma (RB) represents the most common malignant childhood eye tumor worldwide. Several studies indicate that the extracellular matrix (ECM) plays a crucial role in tumor growth and metastasis. Moreover, recent studies indicate that the ECM composition might influence the development of resistance to chemotherapy drugs. The objective of this study was to evaluate possible expression differences in the ECM compartment of the parental human cell lines WERI-RB1 (retinoblastoma 1) and Y79 and their Etoposide resistant subclones via polymerase chain reaction (PCR). Western blot analyses were performed to analyze protein levels. To explore the influence of ECM molecules on RB cell proliferation, death, and cluster formation, WERI-RB1 and resistant WERI-ETOR cells were cultivated on Fibronectin, Laminin, Tenascin-C, and Collagen IV and analyzed via time-lapse video microscopy as well as immunocytochemistry. We revealed a significantly reduced mRNA expression of the proteoglycans <i>Brevican</i>, <i>Neurocan,</i> and <i>Versican</i> in resistant WERI-ETOR compared to sensitive WERI-RB1 cells. Also, for the glycoproteins <i>α1-Laminin</i>, <i>Fibronectin</i>, <i>Tenascin-C,</i> and <i>Tenascin-R</i> as well as <i>Collagen IV</i>, reduced expression levels were observed in WERI-ETOR. Furthermore, a downregulation was detected for the matrix metalloproteinases <i>MMP2</i>, <i>MMP7</i>, <i>MMP9</i>, the tissue-inhibitor of metalloproteinase <i>TIMP2</i>, the Integrin receptor subunits <i>ITGA4</i>, <i>ITGA5</i> and <i>ITGB1,</i> and all <i>receptor protein tyrosine phosphatase β/ζ</i> isoforms. Downregulation of <i>Brevican</i>, <i>Collagen IV</i>, <i>Tenascin-R</i>, <i>MMP2</i>, <i>TIMP2,</i> and <i>ITGA5</i> was also verified in Etoposide resistant Y79 cells compared to sensitive ones. Protein levels of Tenascin-C and MMP-2 were comparable in both WERI cell lines. Interestingly, Fibronectin displayed an apoptosis-inducing effect on WERI-RB1 cells, whereas an anti-apoptotic influence was observed for Tenascin-C. Conversely, proliferation of WERI-ETOR cells was enhanced on Tenascin-C, while an anti-proliferative effect was observed on Fibronectin. In WERI-ETOR, cluster formation was decreased on the substrates Collagen IV, Fibronectin, and Tenascin-C. Collectively, we noted a different ECM mRNA expression and behavior of Etoposide resistant compared to sensitive RB cells. These findings may indicate a key role of ECM components in chemotherapy resistance formation of RB.https://www.mdpi.com/1422-0067/21/12/4322chemotherapy resistanceetoposideextracellular matrixintegrinsretinoblastomaWERI-RB1 |
| spellingShingle | Jacqueline Reinhard Natalie Wagner Miriam M. Krämer Marvin Jarocki Stephanie C. Joachim H. Burkhard Dick Andreas Faissner Vinodh Kakkassery Expression Changes and Impact of the Extracellular Matrix on Etoposide Resistant Human Retinoblastoma Cell Lines International Journal of Molecular Sciences chemotherapy resistance etoposide extracellular matrix integrins retinoblastoma WERI-RB1 |
| title | Expression Changes and Impact of the Extracellular Matrix on Etoposide Resistant Human Retinoblastoma Cell Lines |
| title_full | Expression Changes and Impact of the Extracellular Matrix on Etoposide Resistant Human Retinoblastoma Cell Lines |
| title_fullStr | Expression Changes and Impact of the Extracellular Matrix on Etoposide Resistant Human Retinoblastoma Cell Lines |
| title_full_unstemmed | Expression Changes and Impact of the Extracellular Matrix on Etoposide Resistant Human Retinoblastoma Cell Lines |
| title_short | Expression Changes and Impact of the Extracellular Matrix on Etoposide Resistant Human Retinoblastoma Cell Lines |
| title_sort | expression changes and impact of the extracellular matrix on etoposide resistant human retinoblastoma cell lines |
| topic | chemotherapy resistance etoposide extracellular matrix integrins retinoblastoma WERI-RB1 |
| url | https://www.mdpi.com/1422-0067/21/12/4322 |
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