A Cytokeratin 8 (K8)/Green Fluorescent Protein (GFP) fusion construct was created to better understand the behavior of K8 within cancer cells. This intermediate filament (IF) protein is a member of the cytoskeletal gene family along with actin and tubulin. IF's are normally expressed in a tissue specific and differentiation dependant manner, in which their role is more supportive than essential to the cell. Such roles include rigidity of cellular shape, protein trafficking, cellular locomotion, and cell signaling platforms. K8 mutation, over expression, and aberrant post translational modifications have been observed in various carcinoma cell lines to be the cause of several phenotypes including apoptosis inhibition, drug resistance, transformation, Mallory-Denk body (MDB) formation, localization at the plasma membrane, and secretion of the protein. In order to study these abnormal phenotypes the K8 gene was generated and inserted into the GFP over expression vector. This allowed for the study of K8 within a well defined cervical cancer cell line named Hela. This study intended to provide answers to K8's localization at the plasma membrane in carcinoma cell models while avoiding criticisms to previous immunohistochemical localization studies. A model which exhibits established phenotypes found in the literature was thus created which has the potential to address several paramount questions related to K8's role in supporting the development and progression of cancer. It could also be utilized as an assay for the discovery of K8 filament formation inhibitors, which may prove useful in combination with current chemotherapeutics. The model could also be used to provide weight to diagnostics, such as the Cancer Recognition test, which utilizes antibodies against K8 as biomarkers for malignancy via an Enzyme-Link ImmunoSorbent Assay (ELISA).
| Identifer | oai:union.ndltd.org:ucf.edu/oai:stars.library.ucf.edu:honorstheses1990-2015-1795 |
| Date | 01 January 2008 |
| Creators | Leventhal, Daniel S. |
| Publisher | STARS |
| Source Sets | University of Central Florida |
| Language | English |
| Detected Language | English |
| Type | text |
| Source | HIM 1990-2015 |
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