Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retention
Abstract Background Triple negative breast cancer (TNBC) is an aggressive subtype of breast cancer with limited therapeutic opportunities. Recently, splicing factors have gained attention as potential targets for cancer treatment. Here we systematically evaluated the role of RNA splicing factors in...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2021-03-01
|
| Series: | Journal of Experimental & Clinical Cancer Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13046-021-01863-4 |
| _version_ | 1818675928530681856 |
|---|---|
| author | Esmee Koedoot Eline van Steijn Marjolein Vermeer Román González-Prieto Alfred C. O. Vertegaal John W. M. Martens Sylvia E. Le Dévédec Bob van de Water |
| author_facet | Esmee Koedoot Eline van Steijn Marjolein Vermeer Román González-Prieto Alfred C. O. Vertegaal John W. M. Martens Sylvia E. Le Dévédec Bob van de Water |
| author_sort | Esmee Koedoot |
| collection | DOAJ |
| description | Abstract Background Triple negative breast cancer (TNBC) is an aggressive subtype of breast cancer with limited therapeutic opportunities. Recently, splicing factors have gained attention as potential targets for cancer treatment. Here we systematically evaluated the role of RNA splicing factors in TNBC cell proliferation. Methods In this study, we performed an RNAi screen targeting 244 individual splicing factors to systematically evaluate their role in TNBC cell proliferation. For top candidates, mechanistic insight was gained using amongst others western blot, PCR, FACS, molecular imaging and cloning. Pulldown followed by mass spectrometry were used to determine protein-protein interactions and patient-derived RNA sequencing data was used relate splicing factor expression levels to proliferation markers. Results We identified nine splicing factors, including SNRPD2, SNRPD3 and NHP2L1, of which depletion inhibited proliferation in two TNBC cell lines by deregulation of sister chromatid cohesion (SCC) via increased sororin intron 1 retention and down-regulation of SMC1, MAU2 and ESPL1. Protein-protein interaction analysis of SNRPD2, SNRPD3 and NHP2L1 identified that seven out of the nine identified splicing factors belong to the same spliceosome complex including novel component SUN2 that was also critical for efficient sororin splicing. Finally, sororin transcript levels are highly correlated to various proliferation markers in BC patients. Conclusion We systematically determined splicing factors that control proliferation of breast cancer cells through a mechanism that involves effective sororin splicing and thereby appropriate sister chromatid cohesion. Moreover, we identified SUN2 as an important new spliceosome complex interacting protein that is critical in this process. We anticipate that deregulating sororin levels through targeting of the relevant splicing factors might be a potential strategy to treat TNBC. |
| first_indexed | 2024-12-17T08:35:22Z |
| format | Article |
| id | doaj.art-965c99b87fb3412fb4c7057176e1ddb5 |
| institution | Directory Open Access Journal |
| issn | 1756-9966 |
| language | English |
| last_indexed | 2024-12-17T08:35:22Z |
| publishDate | 2021-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Experimental & Clinical Cancer Research |
| spelling | doaj.art-965c99b87fb3412fb4c7057176e1ddb52022-12-21T21:56:29ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662021-03-0140111710.1186/s13046-021-01863-4Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retentionEsmee Koedoot0Eline van Steijn1Marjolein Vermeer2Román González-Prieto3Alfred C. O. Vertegaal4John W. M. Martens5Sylvia E. Le Dévédec6Bob van de Water7Division of Drug Discovery and Safety, LACDR, Leiden UniversityDivision of Drug Discovery and Safety, LACDR, Leiden UniversityDivision of Drug Discovery and Safety, LACDR, Leiden UniversityDepartment of Molecular Cell Biology, Leiden University Medical CenterDepartment of Molecular Cell Biology, Leiden University Medical CenterDepartment of Medical Oncology and Cancer Genomics Netherlands, Erasmus MC Cancer Institute, Erasmus University Medical CenterDivision of Drug Discovery and Safety, LACDR, Leiden UniversityDivision of Drug Discovery and Safety, LACDR, Leiden UniversityAbstract Background Triple negative breast cancer (TNBC) is an aggressive subtype of breast cancer with limited therapeutic opportunities. Recently, splicing factors have gained attention as potential targets for cancer treatment. Here we systematically evaluated the role of RNA splicing factors in TNBC cell proliferation. Methods In this study, we performed an RNAi screen targeting 244 individual splicing factors to systematically evaluate their role in TNBC cell proliferation. For top candidates, mechanistic insight was gained using amongst others western blot, PCR, FACS, molecular imaging and cloning. Pulldown followed by mass spectrometry were used to determine protein-protein interactions and patient-derived RNA sequencing data was used relate splicing factor expression levels to proliferation markers. Results We identified nine splicing factors, including SNRPD2, SNRPD3 and NHP2L1, of which depletion inhibited proliferation in two TNBC cell lines by deregulation of sister chromatid cohesion (SCC) via increased sororin intron 1 retention and down-regulation of SMC1, MAU2 and ESPL1. Protein-protein interaction analysis of SNRPD2, SNRPD3 and NHP2L1 identified that seven out of the nine identified splicing factors belong to the same spliceosome complex including novel component SUN2 that was also critical for efficient sororin splicing. Finally, sororin transcript levels are highly correlated to various proliferation markers in BC patients. Conclusion We systematically determined splicing factors that control proliferation of breast cancer cells through a mechanism that involves effective sororin splicing and thereby appropriate sister chromatid cohesion. Moreover, we identified SUN2 as an important new spliceosome complex interacting protein that is critical in this process. We anticipate that deregulating sororin levels through targeting of the relevant splicing factors might be a potential strategy to treat TNBC.https://doi.org/10.1186/s13046-021-01863-4Triple-negative breast cancerMitosisSister chromatid cohesionRNA splicingSplicing factorsProliferation |
| spellingShingle | Esmee Koedoot Eline van Steijn Marjolein Vermeer Román González-Prieto Alfred C. O. Vertegaal John W. M. Martens Sylvia E. Le Dévédec Bob van de Water Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retention Journal of Experimental & Clinical Cancer Research Triple-negative breast cancer Mitosis Sister chromatid cohesion RNA splicing Splicing factors Proliferation |
| title | Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retention |
| title_full | Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retention |
| title_fullStr | Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retention |
| title_full_unstemmed | Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retention |
| title_short | Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retention |
| title_sort | splicing factors control triple negative breast cancer cell mitosis through sun2 interaction and sororin intron retention |
| topic | Triple-negative breast cancer Mitosis Sister chromatid cohesion RNA splicing Splicing factors Proliferation |
| url | https://doi.org/10.1186/s13046-021-01863-4 |
| work_keys_str_mv | AT esmeekoedoot splicingfactorscontroltriplenegativebreastcancercellmitosisthroughsun2interactionandsororinintronretention AT elinevansteijn splicingfactorscontroltriplenegativebreastcancercellmitosisthroughsun2interactionandsororinintronretention AT marjoleinvermeer splicingfactorscontroltriplenegativebreastcancercellmitosisthroughsun2interactionandsororinintronretention AT romangonzalezprieto splicingfactorscontroltriplenegativebreastcancercellmitosisthroughsun2interactionandsororinintronretention AT alfredcovertegaal splicingfactorscontroltriplenegativebreastcancercellmitosisthroughsun2interactionandsororinintronretention AT johnwmmartens splicingfactorscontroltriplenegativebreastcancercellmitosisthroughsun2interactionandsororinintronretention AT sylviaeledevedec splicingfactorscontroltriplenegativebreastcancercellmitosisthroughsun2interactionandsororinintronretention AT bobvandewater splicingfactorscontroltriplenegativebreastcancercellmitosisthroughsun2interactionandsororinintronretention |