Investigating the Tumor Suppressive Role of RYBP in U-87 Glioblastoma Cells

Poster Number

9

Submitting Student(s)

Catherine Moorhouse

Session Title

Poster Session 1

College

College of Arts and Sciences

Department

Biology

Abstract

Glioblastoma multiforme (GBM) is the most common and aggressive form of brain cancer. GBM tumors extend threadlike tendrils throughout the brain, which complicates or even eliminates the option of surgical resection. Even with standard chemotherapy and radiation, the median survival rate in adults is only around 14 months. Moreover, GBM displays increased intra- and inter-tumor heterogeneity, which leads to therapeutic resistance and recurrent disease. Thus, there is an urgent need to understand targetable molecular networks that drive GBM progression. Evidence suggests that RING1- and YY1-Binding Protein (RYBP) serves as a tumor suppressor in multiple cancers. RYBP has been recognized as a tumor suppressor in a series of cancers, and in GBM, RYBP expression levels are reduced and inversely correlate with patient prognosis. However, its molecular role in GBM is not well understood. Therefore, we hypothesized that RYBP exerts a tumor suppressive effect in GBM. To test the tumor suppressive effects of RYBP on GBM cells, we generated RYBP-expressing plasmids and co-transfected them into U-87 cells alongside a GFP-expressing reporter plasmid. After 24 hours, we isolated total protein from transfected cells and verified RYBP expression by Western blot. Our data suggest that none of our plasmid clones successfully forced RYBP expression in U-87 cells, so we were unable to measure RYBP’s effects on GBM cell proliferation. In the future, we will need to isolate plasmids from different bacterial clones and sequence them to ensure they do not contain mutations that may inhibit transient manipulation of RYBP expression in GBM cells.

Start Date

15-4-2022 12:00 PM

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Apr 15th, 12:00 PM

Investigating the Tumor Suppressive Role of RYBP in U-87 Glioblastoma Cells

Glioblastoma multiforme (GBM) is the most common and aggressive form of brain cancer. GBM tumors extend threadlike tendrils throughout the brain, which complicates or even eliminates the option of surgical resection. Even with standard chemotherapy and radiation, the median survival rate in adults is only around 14 months. Moreover, GBM displays increased intra- and inter-tumor heterogeneity, which leads to therapeutic resistance and recurrent disease. Thus, there is an urgent need to understand targetable molecular networks that drive GBM progression. Evidence suggests that RING1- and YY1-Binding Protein (RYBP) serves as a tumor suppressor in multiple cancers. RYBP has been recognized as a tumor suppressor in a series of cancers, and in GBM, RYBP expression levels are reduced and inversely correlate with patient prognosis. However, its molecular role in GBM is not well understood. Therefore, we hypothesized that RYBP exerts a tumor suppressive effect in GBM. To test the tumor suppressive effects of RYBP on GBM cells, we generated RYBP-expressing plasmids and co-transfected them into U-87 cells alongside a GFP-expressing reporter plasmid. After 24 hours, we isolated total protein from transfected cells and verified RYBP expression by Western blot. Our data suggest that none of our plasmid clones successfully forced RYBP expression in U-87 cells, so we were unable to measure RYBP’s effects on GBM cell proliferation. In the future, we will need to isolate plasmids from different bacterial clones and sequence them to ensure they do not contain mutations that may inhibit transient manipulation of RYBP expression in GBM cells.