Alpha Omega Alpha Honor Medical Society

2014 Research Abstract

The roles of EMR2 and EMR3 in invasiveness in glioblastoma: evaluation of cellular migration and investigation of downstream pathways

Investigator: Phillip Bonney, University of Oklahoma College of Medicine

Mentor: Michael Sughrue, MD

Introduction: Epidermal growth factor (EGF)-like module containing mucin-like receptors 2 and 3 (EMR2 and EMR3) are seven-span transmembrane G protein coupled receptors that have been studied in immune cells, particularly activated macrophages. Data from The Cancer Genome Atlas (TCGA) have shown that overexpression of these receptors is associated with a poor prognosis in glioblastoma, the most common primary brain cancer in adults. We sought to further investigate the effects of these receptors in glioblastoma.

Methods: Stable overexpressing EMR2 (U251, G55) and EMR3 (U87, U251, G55) lines were created using lentiviral transfection. To assess migration, these cells and LacZ-overexpressing controls were seeded in 3-dimensional microfluidic chambers, with each chamber containing 10 lanes in which cells migrate. Migration velocities were determined by photographing the cells at 30 minute intervals and measuring the distances traveled. The experiment was repeated 5 times to attain statistical power.

Results: Evidence of overexpression was demonstrated through immunofluorescence and Western blot. Increased velocities were observed in U87 cells overexpressing EMR3 compared to LacZ controls (16 μm/hr 1 vs. 10 μm/hr, p < 0.05). No statistical difference was detected between EMR3 and controls in the U251 or G55 lines. No differences were detected between EMR2 and controls in the U251 or G55 lines.

Ongoing Studies: RNA expression analysis is currently being performed on the cell lines in an attempt to characterize the pathways used by EMR2 and EMR3. We have developed a glioblastoma xenograft model through this project, which is currently being used to evaluate for a difference between U87-EMR3 and U87-LacZ in vivo. This experiment will study invasion of tumor cells using immunohistochemistry and survival using Kaplan Meier analysis.

Last modified: 8/14/2015

Updated on August 14, 2015.

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