Wound healing and the management of chronic, non-healing wounds represent a significant burden to the NHS and results in substantial patient morbidity. New methods to further understanding of wound chronicity and potential therapies are needed. This PhD study aims to explore the importance of the SOCS family in the wound healing process, exploring their potential to act as prognostic factors in clinical cohorts and exploring the potential of SOCS-3 and -4 to influence key cellular traits linked to the healing process. Detection of SOCS family members within a clinical healing/non-healing cohort highlighted significant elevations of gene expression of SOCS-3 and -4 in non-healing chronic wounds compared to healing chronic wounds, though this trend was no as obvious in a smaller cohort of IHC stained samples. However, the IHC stained samples potentially indicated that SOCS-3 protein relocalisation, from wound edge to distal wound area, was evident in the non-healing chronic wound. Subsequently, SOCS-3 expression and SOCS-4 knockdown lines were generated in human (HaCaT) keratinocytes and (HECV) endothelial cells to explore the functional significance of these molecules at a cellular level. In summary, SOCS-4 knockdown decreased the migration of both HaCaT and HECV cells on plastic cultureware, whereas downregulation of SOCS-4 only reduced the adhesion and tubule formation of HECV cells on matrigel matrix. SOCS-3 expression attenuated the proliferation of HaCaT cells but had no effect on HECV cells. In addition, SOCS-3 upregulation solely improved the adhesion and tubule formation of HECV cells on matrigel matrix. Moreover, the regulatory role of SOCS-3 and SOCS-4 in the migration and adhesion of HaCaT and HECV are likely to be substratum matrix dependent. Finally, to explore potential mechanisms of action for SOCS-4 role in HaCaT cell line, a protein microarray was utilised to highlight a number of potential key pathways. Based on the results from protein microarray and wound healing assay as well as evidences from literature review, four potential mechanisms (FAK/Src/p130cas, HBEGF/EGFR, VEGF/VEGFR-2, IGF-1/IGF-1R/PI3K) of SOCS-4 relulating keratinocyte migration was highlighted for further validation, and the expression profile of FAK Y397 was varified by western blotting. Hence, SOCS-3 and SOCS-4 may regulate keratinocyte and endothelial cell behaviour, and potentially have effect on the wound healing process.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:709612 |
| Date | January 2017 |
| Creators | Feng, Yi |
| Publisher | Cardiff University |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://orca.cf.ac.uk/99327/ |
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