Investigating the initial molecular interactions between hepatocyte- like cells with the extracellularmatrix and hepatic sinusoidal endothelial cells and The loss of LAR inhibits the activation of the PDGF receptor by NHERF2 via the PKC δ pathway

Blakeway, Daniel William (2013). Investigating the initial molecular interactions between hepatocyte- like cells with the extracellularmatrix and hepatic sinusoidal endothelial cells and The loss of LAR inhibits the activation of the PDGF receptor by NHERF2 via the PKC δ pathway. University of Birmingham. M.Res.

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Abstract

Project 1: Interactions between ES-HLCs with the ECM and HSEC. Flow cytometry was carried out on various types of HSEC to quantify the level of the integrins being expressed. Static adhesion assays were performed on ECM proteins to give a measure of cell adhesion. Flow cytometry showed integrin-β1 and BCAM were highly expressed. ES-HLCs showed preferential binding to Laminin, adherence was significantly reduced when the cells were treated with β1 and BCAM blocking antibodies; ES-HLC adherence was reduced when laminin was pre-treated with recombinant BCAM/Fs.

Project 2: The role of leukocyte common antigen-related receptor (LAR-RPTP), in the regulation of the PDGF receptor. PKC δ an enzyme protein, showed to regulate the PDGF; through inhibiting tyrosine phosphorylation. Structural proteins NHERF1 and NHERF2 showed to bind to the PDGF, causing receptor dimerization. siRNA knockdown of PKC δ, NHERF1 and NHERF2 was performed in the absence of LAR, and analysed through western blot. Data shows in the absence of PKC δ and NHERF-2, PDGFR activity increased; in the absence of NHERF1 the activity was inhibited. Results show that LAR regulates PDGFR signaling through activation of the PDGF-β, reducing the activation of PKC δ inhibiting the PDGF-β activation whilst dissociates NHERF1 and promoting NHERF2 binding.

Type of Work:

Thesis (Masters by Research > M.Res.)

Award Type:

Masters by Research > M.Res.

Supervisor(s):