The role of plectin in the regulation of carcinoma cell invasion

Abstract

The research presented in this thesis endeavours to understand the role of the plakin family of cytoskeletal linker proteins in the migration and invasive potential of epithelial carcinomas. This study focuses on plectin, a plakin family member that has previously been implicated with a role in tissue integrity of the skin and muscle. I investigated the complex gene organisation of the alternative first exons in human plectin, leading to the discovery of a further novel isoform plectin-lk. A panel of colon and breast carcinoma cell lines that vary in their differentiation and metastatic potential were used to investigate the expression of the isoforms at the mRNA and protein levels, showing plectin to be expressed at higher levels in the more invasive cell types. The subcellular localisation of the alternatively spliced plectin isoforms was investigated using green fluorescent protein (GFP) and polyclonal antibodies, revealing isoform specific targeting to different actin structures. Ablation of plectin or vimentin (a major intermediate filament protein of mesenchymal cells that interacts with plectin), by small interfering RNAs suggest these proteins are able to modulate invasion, migration and attachment of the epithelial carcinoma cells. Further investigation into the novel isoform plectin-lk reveals a role in the formation of podosome like adhesion structures in a Rho kinase dependant manner that facilitate migration in the SW480 colon carcinoma cells. The above findings are novel and contribute to the understanding of migration and invasion of cancer cells. Furthermore, this understanding could provide novel targets of cancer cell metastasis.