The Role of DNA Methylation in RYBP Silencing in Glioblastoma Cells

Submitting Student(s)

Michelle Aguilar-Gaspar

Session Title

Poster Session 1

Faculty Mentor

Daniel Stovall, Ph.D.

College

College of Arts and Sciences

Department

Biology

Abstract

Glioblastoma multiforme (GBM) is a malignant type of central nervous system tumor that is highly invasive, which makes surgical resection difficult and contributes to disease recurrence. Because GBM is the most lethal and common tumor of the brain and spinal cord, it has a median survival of less than 15 months. This median survival highlights the need for more effective and targeted therapeutic options. GBM tumors frequently downregulate RYBP expression, compared to healthy brain tissue. In many types of solid tumors, RYBP functions as a tumor suppressor gene and restricts cancer growth. With this in mind, it is possible that the reactivation of RYBP in GBM may exert similar growth-inhibiting effects; however, the mechanism by which RYBP is silenced remains unclear. The aim of this study was to investigate whether DNA methylation in the RYBP promoter contributed to the downregulation of RYBP in GBM cells. We inhibited DNA methylation by treating U118 and T98 GBM cell lines with 5-aza-2’deoxycytidine (5-aza) or DMSO (vehicle control). Seventy-two hours after drug administration, total RNA was isolated, quantified, and used for reverse-transcription-qPCR to verify RYBP expression. Our findings show RYBP levels increased in a dose-dependent manner following 5-aza treatment. However, these experiments need to be repeated to determine statistical significance. At present, it is unclear whether the reactivation of RYBP by 5-aza is due to direct or indirect effects. Therefore, a future goal of this project is to isolate genomic DNA and examine the methylation status of the RYBP gene promoter.

Previously Presented/Performed?

Winthrop University Showcase of Undergraduate Research and Creative Endeavors, Rock Hill, SC, April 2023.

Type of Presentation

Poster presentation

Grant Support?

Supported by SC-INBRE DRP 5P20GM103499-21 and the Winthrop University Research Council

Start Date

15-4-2023 12:00 PM

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Apr 15th, 12:00 PM

The Role of DNA Methylation in RYBP Silencing in Glioblastoma Cells

Glioblastoma multiforme (GBM) is a malignant type of central nervous system tumor that is highly invasive, which makes surgical resection difficult and contributes to disease recurrence. Because GBM is the most lethal and common tumor of the brain and spinal cord, it has a median survival of less than 15 months. This median survival highlights the need for more effective and targeted therapeutic options. GBM tumors frequently downregulate RYBP expression, compared to healthy brain tissue. In many types of solid tumors, RYBP functions as a tumor suppressor gene and restricts cancer growth. With this in mind, it is possible that the reactivation of RYBP in GBM may exert similar growth-inhibiting effects; however, the mechanism by which RYBP is silenced remains unclear. The aim of this study was to investigate whether DNA methylation in the RYBP promoter contributed to the downregulation of RYBP in GBM cells. We inhibited DNA methylation by treating U118 and T98 GBM cell lines with 5-aza-2’deoxycytidine (5-aza) or DMSO (vehicle control). Seventy-two hours after drug administration, total RNA was isolated, quantified, and used for reverse-transcription-qPCR to verify RYBP expression. Our findings show RYBP levels increased in a dose-dependent manner following 5-aza treatment. However, these experiments need to be repeated to determine statistical significance. At present, it is unclear whether the reactivation of RYBP by 5-aza is due to direct or indirect effects. Therefore, a future goal of this project is to isolate genomic DNA and examine the methylation status of the RYBP gene promoter.