P2X7 is a ligand-gated ion channel activated by the extracellular nucleotide ATP. Although P2X7 was originally identified as a cytolytic receptor on immune cells, activation of P2X7 can also contribute to increased cell proliferation in cancer cells. Studies have shown that selective antagonists of P2X7 can slow cancer cell growth in vitro and in vivo. A number of splice variants have been identified for P2X7 and C-terminal truncated variants such as P2X7B are implicated in this proliferative effect which may be linked to cellular metabolic reprogramming. This project will explore the expression of P2X7 splice variants in various cancers (e.g. melanoma, breast, prostate) using well documented cell lines in 2D and 3D cell culture models and the correlation with aggressiveness. In addition to splice variants which may change large regions of the ion channel structure, many point mutations exist in P2X7 due to inheritance of single nucleotide polymorphisms in the P2RX7 gene. We will genotype the studied cancer cell lines for non-synonymous SNPs and investigate the impact of P2RX7 haplotypes on cancer cell biology. We will determine if there is correlation between particular SNPs or splice variants and cancer aggressiveness and potentially whether this information could be used as a biomarker for prognosis.
Techniques: Cell culture, viability, cell metabolism, migration and invasion assays will be performed and the student will also be trained in several molecular biology techniques including quantitative PCR, western blotting, flow cytometry, and plate-reader fluorescence assays.