A novel non-sulphamoylated 2-methoxyestradiol derivative causes detachment of breast cancer cells by rapid disassembly of focal adhesions
Abstract Background 2-Methoxyestradiol (2ME2) is an estradiol metabolite with well documented antiproliferative properties in many cancer cell lines. However, it is rapidly metabolised in vivo which limits its clinical application. Therefore, more stable derivatives with potentially improved clinica...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2018-11-01
|
| Series: | Cancer Cell International |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12935-018-0688-7 |
| _version_ | 1819067981721763840 |
|---|---|
| author | Mandie Botes Tamarin Jurgens Zohreh Riahi Michelle Visagie Rustelle Janse van Vuuren Anna Margaretha Joubert Iman van den Bout |
| author_facet | Mandie Botes Tamarin Jurgens Zohreh Riahi Michelle Visagie Rustelle Janse van Vuuren Anna Margaretha Joubert Iman van den Bout |
| author_sort | Mandie Botes |
| collection | DOAJ |
| description | Abstract Background 2-Methoxyestradiol (2ME2) is an estradiol metabolite with well documented antiproliferative properties in many cancer cell lines. However, it is rapidly metabolised in vivo which limits its clinical application. Therefore, more stable derivatives with potentially improved clinical features have been designed by our group. Here we describe an estrone-like derivative of 2ME2, namely EE-15-one, that unlike other derivatives which induce cell cycle arrest, induces a rapid loss of cell–substrate adhesion through the inactivation and disassembly of focal adhesions. Methods To assess the effect of 2-ethyl-estra-1,3,5 (10),15-tetraen-3-ol-17-one (EE-15-one) on breast cancer cell lines, cell survival was quantified. The effect of EE-15-one on cell attachment was assessed by measuring cell adhesion and cell rounding via light microscopy. Effects on focal adhesion dynamics and actin cytoskeleton organisation were visualised by immunofluorescence while focal adhesion signalling was assessed by western blot. Cell death was quantified using a lactate dehydrogenase activity (LDH) assay. To investigate specificity towards cell–substrate over cell–cell contact inhibition, EE-15-one effects on 3D cell cultures were assessed. Results Cell survival assays show an almost complete loss of cells within 24 h of EE-15-one exposure in contrast to published sulphamoylated 2ME2 derivatives. Cell loss is linked to rapid detachment and adhesion inhibition. Focal adhesion size and number are rapidly diminished while actin fibres became severed and disappeared within 2 h post exposure. These changes were not due to cell necrosis as LDH activity only slightly increased after 24 h. Cells grown in cell–cell adhesion dependent spheroids did not respond to EE-15-one exposure suggesting that EE-15-one specifically inhibits cell–substrate adhesions but not cell–cell adhesions and does not directly impact the actin cytoskeleton. Conclusion We show that a novel 2ME2 derivative, EE-15-one, induces rapid loss of focal adhesion function leading to cell–substrate detachment through interference with integrin-based cell–substrate adhesions, but not cadherin dependent cell–cell adhesions. Therefore, EE-15-one is the first 2ME2 derivative that has an alternative mode of action to the antimitotic activity of 2ME2. As such EE-15-one shows potential as a lead compound for further development as an inhibitor of cell–substrate adhesion which is essential for metastatic dissemination. |
| first_indexed | 2024-12-21T16:26:53Z |
| format | Article |
| id | doaj.art-0aaf460e1e65438299e76fc51d74ddac |
| institution | Directory Open Access Journal |
| issn | 1475-2867 |
| language | English |
| last_indexed | 2024-12-21T16:26:53Z |
| publishDate | 2018-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Cancer Cell International |
| spelling | doaj.art-0aaf460e1e65438299e76fc51d74ddac2022-12-21T18:57:26ZengBMCCancer Cell International1475-28672018-11-0118111210.1186/s12935-018-0688-7A novel non-sulphamoylated 2-methoxyestradiol derivative causes detachment of breast cancer cells by rapid disassembly of focal adhesionsMandie Botes0Tamarin Jurgens1Zohreh Riahi2Michelle Visagie3Rustelle Janse van Vuuren4Anna Margaretha Joubert5Iman van den Bout6Department of Physiology, University of PretoriaDepartment of Physiology, University of PretoriaDepartment of Physiology, University of PretoriaDepartment of Physiology, University of PretoriaDepartment of Physiology, University of PretoriaDepartment of Physiology, University of PretoriaDepartment of Physiology, University of PretoriaAbstract Background 2-Methoxyestradiol (2ME2) is an estradiol metabolite with well documented antiproliferative properties in many cancer cell lines. However, it is rapidly metabolised in vivo which limits its clinical application. Therefore, more stable derivatives with potentially improved clinical features have been designed by our group. Here we describe an estrone-like derivative of 2ME2, namely EE-15-one, that unlike other derivatives which induce cell cycle arrest, induces a rapid loss of cell–substrate adhesion through the inactivation and disassembly of focal adhesions. Methods To assess the effect of 2-ethyl-estra-1,3,5 (10),15-tetraen-3-ol-17-one (EE-15-one) on breast cancer cell lines, cell survival was quantified. The effect of EE-15-one on cell attachment was assessed by measuring cell adhesion and cell rounding via light microscopy. Effects on focal adhesion dynamics and actin cytoskeleton organisation were visualised by immunofluorescence while focal adhesion signalling was assessed by western blot. Cell death was quantified using a lactate dehydrogenase activity (LDH) assay. To investigate specificity towards cell–substrate over cell–cell contact inhibition, EE-15-one effects on 3D cell cultures were assessed. Results Cell survival assays show an almost complete loss of cells within 24 h of EE-15-one exposure in contrast to published sulphamoylated 2ME2 derivatives. Cell loss is linked to rapid detachment and adhesion inhibition. Focal adhesion size and number are rapidly diminished while actin fibres became severed and disappeared within 2 h post exposure. These changes were not due to cell necrosis as LDH activity only slightly increased after 24 h. Cells grown in cell–cell adhesion dependent spheroids did not respond to EE-15-one exposure suggesting that EE-15-one specifically inhibits cell–substrate adhesions but not cell–cell adhesions and does not directly impact the actin cytoskeleton. Conclusion We show that a novel 2ME2 derivative, EE-15-one, induces rapid loss of focal adhesion function leading to cell–substrate detachment through interference with integrin-based cell–substrate adhesions, but not cadherin dependent cell–cell adhesions. Therefore, EE-15-one is the first 2ME2 derivative that has an alternative mode of action to the antimitotic activity of 2ME2. As such EE-15-one shows potential as a lead compound for further development as an inhibitor of cell–substrate adhesion which is essential for metastatic dissemination.http://link.springer.com/article/10.1186/s12935-018-0688-7Breast cancer2-MethoxyestradiolCell detachmentFocal adhesion |
| spellingShingle | Mandie Botes Tamarin Jurgens Zohreh Riahi Michelle Visagie Rustelle Janse van Vuuren Anna Margaretha Joubert Iman van den Bout A novel non-sulphamoylated 2-methoxyestradiol derivative causes detachment of breast cancer cells by rapid disassembly of focal adhesions Cancer Cell International Breast cancer 2-Methoxyestradiol Cell detachment Focal adhesion |
| title | A novel non-sulphamoylated 2-methoxyestradiol derivative causes detachment of breast cancer cells by rapid disassembly of focal adhesions |
| title_full | A novel non-sulphamoylated 2-methoxyestradiol derivative causes detachment of breast cancer cells by rapid disassembly of focal adhesions |
| title_fullStr | A novel non-sulphamoylated 2-methoxyestradiol derivative causes detachment of breast cancer cells by rapid disassembly of focal adhesions |
| title_full_unstemmed | A novel non-sulphamoylated 2-methoxyestradiol derivative causes detachment of breast cancer cells by rapid disassembly of focal adhesions |
| title_short | A novel non-sulphamoylated 2-methoxyestradiol derivative causes detachment of breast cancer cells by rapid disassembly of focal adhesions |
| title_sort | novel non sulphamoylated 2 methoxyestradiol derivative causes detachment of breast cancer cells by rapid disassembly of focal adhesions |
| topic | Breast cancer 2-Methoxyestradiol Cell detachment Focal adhesion |
| url | http://link.springer.com/article/10.1186/s12935-018-0688-7 |
| work_keys_str_mv | AT mandiebotes anovelnonsulphamoylated2methoxyestradiolderivativecausesdetachmentofbreastcancercellsbyrapiddisassemblyoffocaladhesions AT tamarinjurgens anovelnonsulphamoylated2methoxyestradiolderivativecausesdetachmentofbreastcancercellsbyrapiddisassemblyoffocaladhesions AT zohrehriahi anovelnonsulphamoylated2methoxyestradiolderivativecausesdetachmentofbreastcancercellsbyrapiddisassemblyoffocaladhesions AT michellevisagie anovelnonsulphamoylated2methoxyestradiolderivativecausesdetachmentofbreastcancercellsbyrapiddisassemblyoffocaladhesions AT rustellejansevanvuuren anovelnonsulphamoylated2methoxyestradiolderivativecausesdetachmentofbreastcancercellsbyrapiddisassemblyoffocaladhesions AT annamargarethajoubert anovelnonsulphamoylated2methoxyestradiolderivativecausesdetachmentofbreastcancercellsbyrapiddisassemblyoffocaladhesions AT imanvandenbout anovelnonsulphamoylated2methoxyestradiolderivativecausesdetachmentofbreastcancercellsbyrapiddisassemblyoffocaladhesions AT mandiebotes novelnonsulphamoylated2methoxyestradiolderivativecausesdetachmentofbreastcancercellsbyrapiddisassemblyoffocaladhesions AT tamarinjurgens novelnonsulphamoylated2methoxyestradiolderivativecausesdetachmentofbreastcancercellsbyrapiddisassemblyoffocaladhesions AT zohrehriahi novelnonsulphamoylated2methoxyestradiolderivativecausesdetachmentofbreastcancercellsbyrapiddisassemblyoffocaladhesions AT michellevisagie novelnonsulphamoylated2methoxyestradiolderivativecausesdetachmentofbreastcancercellsbyrapiddisassemblyoffocaladhesions AT rustellejansevanvuuren novelnonsulphamoylated2methoxyestradiolderivativecausesdetachmentofbreastcancercellsbyrapiddisassemblyoffocaladhesions AT annamargarethajoubert novelnonsulphamoylated2methoxyestradiolderivativecausesdetachmentofbreastcancercellsbyrapiddisassemblyoffocaladhesions AT imanvandenbout novelnonsulphamoylated2methoxyestradiolderivativecausesdetachmentofbreastcancercellsbyrapiddisassemblyoffocaladhesions |