Title

An Investigation Into the Effects of VAMP3 on EGFR In Glioblastoma Cells

School Name

South Carolina Governor's School for Science & Mathematics

Grade Level

12th Grade

Presentation Topic

Cell and Molecular Biology

Presentation Type

Mentored

Abstract

Vesicle associated membrane protein 3 (VAMP3) is a protein associated with v-snare structure formation and with endosome trafficking. Through this association, we believe that VAMP3 may play a role in the trafficking of the epidermal growth factor receptor (EGFR) which is a transmembrane protein that has been shown to trigger signaling pathways resulting in the hallmarks of cancer in glioblastoma cells. To investigate the effect of VAMP3 on EGFR, we established a stable VAMP3 knockdown model. Using this model, we ran a recycling assay and used flow cytometry and immunofluorescence data to analyze the levels of the two proteins. We also used a bimolecular fluorescence complementation (BiFC) assay to analyze the interactions between VAMP3, EGFR, and epithelial membrane protein 3 (EMP3) another protein associated with trafficking. We found increased total EGFR levels in our VAMP3 knockdown cells but found no effect on the surface expression of EGFR in the knockdown cells compared to the control cells. We also found evidence of an interaction between EGFR and VAMP3 and confirmed an interaction between EGFR and EMP3. However, we found evidence to support that these three proteins do not work within a complex with each other. Based on these we believe that VAMP3 may play a role in EGFR trafficking but not in the way that we originally anticipated. Further experimentation will shed more light on the interactions between these three proteins. Through further experimentation, we could discover more about the protein interactions that lead to tumor formation.

Location

Furman Hall 107

Start Date

3-28-2020 12:00 PM

Presentation Format

Oral and Written

Group Project

No

COinS
 
Mar 28th, 12:00 PM

An Investigation Into the Effects of VAMP3 on EGFR In Glioblastoma Cells

Furman Hall 107

Vesicle associated membrane protein 3 (VAMP3) is a protein associated with v-snare structure formation and with endosome trafficking. Through this association, we believe that VAMP3 may play a role in the trafficking of the epidermal growth factor receptor (EGFR) which is a transmembrane protein that has been shown to trigger signaling pathways resulting in the hallmarks of cancer in glioblastoma cells. To investigate the effect of VAMP3 on EGFR, we established a stable VAMP3 knockdown model. Using this model, we ran a recycling assay and used flow cytometry and immunofluorescence data to analyze the levels of the two proteins. We also used a bimolecular fluorescence complementation (BiFC) assay to analyze the interactions between VAMP3, EGFR, and epithelial membrane protein 3 (EMP3) another protein associated with trafficking. We found increased total EGFR levels in our VAMP3 knockdown cells but found no effect on the surface expression of EGFR in the knockdown cells compared to the control cells. We also found evidence of an interaction between EGFR and VAMP3 and confirmed an interaction between EGFR and EMP3. However, we found evidence to support that these three proteins do not work within a complex with each other. Based on these we believe that VAMP3 may play a role in EGFR trafficking but not in the way that we originally anticipated. Further experimentation will shed more light on the interactions between these three proteins. Through further experimentation, we could discover more about the protein interactions that lead to tumor formation.