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Prostaglandins are bioactive lipids involved in many physiological functions such as maintenance of the cardiovascular, immune, renal, and central nervous systems. They also play a role in certain diseases like arthritis, cancer, and Alzheimer’s. Cyclooxygenase-2 (COX-2) is the enzyme that catalyzes the initial rate-limiting step in the pathway that converts arachidonic acid to prostaglandins. COX-2 exists as two glycoforms with the molecular weights of 72 and 74 kDa, the latter resulting from the addition of a high mannose chain to the Asn580 residue ~50% of the time. The over-expression of COX-2 is believed to be linked to cancer progression and specifically appears to promote the metastatic phenotype. The objective of this study is to determine the effect of the variable glycosylation of COX-2 at Asn580 on the migratory and invasive potential of cells. COS-1 cells and the breast cancer cell line MCF7 were first transfected with either the wild type or Asn580-mutant human COX-2 gene. Boyden chambers were used to determine the ability of transfected cells to migrate through the membrane, approximately 5x104 cells were plated onto the chambers, and cells were incubated for 16-18 h. Cells were then fixed, stained, visualized and counted. In a previous study, our lab showed that COS-1 cells transfected with the Asn580-mutant COX-2 gene migrated faster through the membrane. In this current study, COS-1 cells transfected with the Asn580-mutant COX-2 gene also had a greater invasive potential; however, MCF7 cells transfected with the wild-type human COX-2 gene migrated faster and also had a greater tendency to invade. The results indicate that the ability of this additional or the lack of glycosylation of COX-2 at Asn580 to either enhance or inhibit the migratory and invasive potential of cells depends greatly on cell type. To confirm this, future studies will be carried out to determine the effect of COX-2 glycosylation on the invasive and the migratory potential of PC-3 and T-47D cancer cell lines.
Natural Sciences and Mathematics
Annual Meeting of the American Society for Cell Biology
Algaly, Fahad; Hand, Julia; and Sevigny, Mary B., "Glycosylation of cyclooxygenase-2 (COX-2) influences the migratory and invasive potential of cells" (2014). Natural Sciences and Mathematics | Faculty Research Posters. 2.