Most human genes produce multiple mRNA isoforms through alternative splicing. However, the biological relevance of most splice variants remains unclear. In this study, we evaluated the functional impact of alternative splicing in cancer cells. We modulated the splicing pattern of 41 cancer-associated splicing events and scored the effects on cell growth, viability and apoptosis, identifying three isoforms essential for cell survival. Specifically, changing the splicing pattern of the spleen tyrosine kinase gene (SYK) impaired cell-cycle progression and anchorage-independent growth. Notably, exposure of cancer cells to epithelial growth factor modulated the SYK splicing pattern to promote the pro-survival isoform that is associated with cancer tissues in vivo. The data suggest that splicing of selected genes is specifically modified during tumor development to allow the expression of isoforms that promote cancer cell survival.
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We are indebted to C. Rancourt (Département de Microbiologie, Université de Sherbrooke) for providing cell lines and help in the initial setup phase of the project and to B. Lamontagne and L. Bergeron Jr. for assay setup and help with troubleshooting. We thank the Réseau de Recherche sur le Cancer (Fonds de la Recherche en Santé du Québec; FRSQ) tissue bank for ovarian tumor tissues and L. Volkov for help with flow cytometry. This work was funded by Genome Canada/Génome Québec and National Cancer Institute of Canada grant no. 700529. B.C. is the Canada Research Chair in Functional Genomics. J.-P.P. is the Canada Research Chair on Genomics and Catalytic RNA. S.A.E. is a Chercheur National of the FRSQ.
The authors declare no competing financial interests.
Supplementary Text and Figures
Supplementary Figures 1–9, Supplementary Tables 1 and 3, and Supplementary Methods (PDF 3282 kb)
Supplementary Table 2
Primary FASE screen results. The results of the primary screen are shown, and the numbers of ISIs that are above the Z-score cut-off (>3) are indicated for each gene and each assay separately. Only ASEs with more than two positive ISIs per assay were retained for the validation phase (phase 2). (XLS 41 kb)
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Prinos, P., Garneau, D., Lucier, JF. et al. Alternative splicing of SYK regulates mitosis and cell survival. Nat Struct Mol Biol 18, 673–679 (2011). https://doi.org/10.1038/nsmb.2040
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