The potential of CLIC proteins as biomarkers for the detection and screening of ovarian cancer
UTS - University of Technology Sydney.
Exosomes (30-100 nm) are extracellular vesicles released by a diverse number of cells, including cancerous body cells. They carry molecular constituents from their cells of origin, principally proteins and RNA (including mRNA and miRNA). It is expected that part of this exosomal cargo would be membrane proteins, including ion channels such as CLICs. When exosomes fuse with the membranes of their target cells, changes in the membrane conductance are expected as their cargo of spontaneously inserting CLIC proteins enter the target cell membrane. This project aims to investigate the profile of such conductance changes using impedance spectroscopy and tethered membranes, and use this knowledge to develop a diagnostic screening platform for cancers that overexpress CLICs. For this study, two ovarian serous adenocarcinoma cell lines were used. As a first step, exosomes secreted by these cells were isolated using a series of ultracentrifugation. The nature of the exosomes particles needs to be verified by identifying the presence of the exosome surface marker CD9, as well as performing NTA (Nanoparticle Tracking Analysis) to measure the effective size of the isolated particles. Western Blot is used to investigate the presence of CLIC proteins in the microvesicles secreted by these ovarian cancer cell lines. For the moment, results from these preliminary steps will be presented.