The Ste20‐like kinase (SLK) has been shown to be expressed in all cell lines and tissues. Previous studies using siRNA and dominant negative approaches have established that SLK plays important roles in cell growth, cytoskeletal dynamics and cell migration. However, the SLK-dependent signaling mechanisms have yet to be elucidated. To further investigate the role of SLK in those processes, we have assessed the effect of a genetic deletion of SLK on cell growth, apoptosis and cell motility. Conditional SLK-floxed fibroblasts were derived and the SLK gene was inactivated in established cell lines. Using cell counts and flow cytometry, we show that SLK deletion does not affect cell growth or progression through the cell cycle. Similarly, Boyden chamber migration assay showed that SLK deletion did not affect cell motility. Using a scratch wound assays and immunofluorescence, we assessed the localization of cytoskeletal protein during migration. Our data show that Paxillin and FAK are still recruited to the leading edge of migrating cells in the absence of SLK. However, nocodazole release studies show that SLK-deficient cells have a faster focal adhesion turnover rate. As SLK has been shown to play a role in apoptosis, we tested the effect of SLK deletion on cell death. Using multiple apoptotic triggers, we show that SLK deletion does not affect apoptosis in fibroblasts and that signaling downstream of those triggers is unchanged. Overall, in contrast to siRNA studies, our data show that the genetic deletion of SLK doesn’t affect cell growth, apoptosis or cell migration. This is likely due to the activation of compensatory mechanisms, bypassing the requirements for SLK.
| Identifer | oai:union.ndltd.org:uottawa.ca/oai:ruor.uottawa.ca:10393/39537 |
| Date | 22 August 2019 |
| Creators | Cunha, Clinton Royce |
| Contributors | Sabourin, Luc A. |
| Publisher | Université d'Ottawa / University of Ottawa |
| Source Sets | Université d’Ottawa |
| Language | English |
| Detected Language | English |
| Type | Thesis |
| Format | application/pdf |
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