Synergistic Effect of the Epithelial Membrane Protein 3 (EMP3) Knockdown with Inhibitors of Tyrosine Kinases on Glioblastoma Cell Death

Peter HoegyFollow

School Name

South Carolina Governor's School for Science and Mathematics

Grade Level

12th Grade

Presentation Topic

Cell and Molecular Biology

Presentation Type

Mentored

Abstract

Glioblastoma (GBM), an aggressive form of brain tumor, contains two deregulated tyrosine kinase proteins, c-Met (Mesenchymal-epithelial transition factor receptor) and EGFR (Epithelial Growth Factor Receptor), causing aggressive proliferation rates. Drugs targeting these proteins show very little promise. However, Epithelial Membrane Protein 3 (EMP3) may play a role in these protein pathways and therefore may prove to be a viable target for GBM. The aim of this research is to explore the synergistic effect between EMP3 gene knockdown (KD) along with c-Met/EGFR inhibitors in GBM cells. Four second generation cell lines were generated from NCH 1425: a) scrRNA + DMSO (SP790); b) scrRNA + DOX (SP790); c) shRNA + DMSO (SP1764); and d) shRNA + DOX (SP1764). Cell lines a-c served as negative controls that did not undergo EMP3 KD while line d experienced KD after induction with antibiotic doxycycline (DOX). These cell lines were then either treated with c-Met inhibitor bozitinib (0-50uM) or EGFR inhibitor osimertinib (0-10uM). The results indicate that there is a general decrease in cell viability with both drugs, regardless of the presence or absence of EMP3. Cell line d+osimertinib shows minimal drop in viability while cell line d+bozitinib shows no drop in viability compared to the control groups. These results could be due to the fact that there was only a 30% EMP3 KD in cell line d, as shown by western blot analysis. In order to understand the synergistic effect, it is imperative that cell line d has nearly a 100% EMP3 KD.

Location

RITA 271

Start Date

3-23-2024 10:15 AM

Presentation Format

Oral Only

Group Project

No

COinS
 
Mar 23rd, 10:15 AM

Synergistic Effect of the Epithelial Membrane Protein 3 (EMP3) Knockdown with Inhibitors of Tyrosine Kinases on Glioblastoma Cell Death

RITA 271

Glioblastoma (GBM), an aggressive form of brain tumor, contains two deregulated tyrosine kinase proteins, c-Met (Mesenchymal-epithelial transition factor receptor) and EGFR (Epithelial Growth Factor Receptor), causing aggressive proliferation rates. Drugs targeting these proteins show very little promise. However, Epithelial Membrane Protein 3 (EMP3) may play a role in these protein pathways and therefore may prove to be a viable target for GBM. The aim of this research is to explore the synergistic effect between EMP3 gene knockdown (KD) along with c-Met/EGFR inhibitors in GBM cells. Four second generation cell lines were generated from NCH 1425: a) scrRNA + DMSO (SP790); b) scrRNA + DOX (SP790); c) shRNA + DMSO (SP1764); and d) shRNA + DOX (SP1764). Cell lines a-c served as negative controls that did not undergo EMP3 KD while line d experienced KD after induction with antibiotic doxycycline (DOX). These cell lines were then either treated with c-Met inhibitor bozitinib (0-50uM) or EGFR inhibitor osimertinib (0-10uM). The results indicate that there is a general decrease in cell viability with both drugs, regardless of the presence or absence of EMP3. Cell line d+osimertinib shows minimal drop in viability while cell line d+bozitinib shows no drop in viability compared to the control groups. These results could be due to the fact that there was only a 30% EMP3 KD in cell line d, as shown by western blot analysis. In order to understand the synergistic effect, it is imperative that cell line d has nearly a 100% EMP3 KD.