Quantification of Cells with Modifications Relating to the RB1 Pathway
Session Number
Project ID: MEDH 39
Advisor(s)
Elizaveta Benevolenskaya, University of Illinois Cancer Center
Discipline
Medical and Health Sciences
Start Date
17-4-2024 9:40 AM
End Date
17-4-2024 9:55 AM
Abstract
The retinoblastoma tumor suppressor (RB1) is a vital tumor suppressor gene. It prevents the cell from transitioning to the S phase from G phase by inhibiting E2F activity which limits cell proliferation and facilitates a stable exit from the cell cycle. Inactivation of RB1 thus allows for the expression of genes necessary for the cell cycle to progress and results in the production of proteins and DNA. RB1 also regulates KDM5A which is a direct repressor of metabolic regulatory genes. Therefore a lack of RB1 causes dysregulation of KDM5A which can lead to downregulation of H3K4me3 levels, effectively silencing the metabolic genes. The EGFR pathway leads to the activation of CDK4/6, which inactivates the RB1 tumor suppression by initiating the phosphorylation of RB1. We raised multiple cell lines which were modified for the presence of RB1 and EGFR TKIs, then added ki67 (a marker for proteins) and EdU (a marker for DNA). By using Zeiss Microscopy Software to quantify the cell images, this project gave insight into the correlation between the expression of KDM5A and E2F within the RB1 pathway.
Quantification of Cells with Modifications Relating to the RB1 Pathway
The retinoblastoma tumor suppressor (RB1) is a vital tumor suppressor gene. It prevents the cell from transitioning to the S phase from G phase by inhibiting E2F activity which limits cell proliferation and facilitates a stable exit from the cell cycle. Inactivation of RB1 thus allows for the expression of genes necessary for the cell cycle to progress and results in the production of proteins and DNA. RB1 also regulates KDM5A which is a direct repressor of metabolic regulatory genes. Therefore a lack of RB1 causes dysregulation of KDM5A which can lead to downregulation of H3K4me3 levels, effectively silencing the metabolic genes. The EGFR pathway leads to the activation of CDK4/6, which inactivates the RB1 tumor suppression by initiating the phosphorylation of RB1. We raised multiple cell lines which were modified for the presence of RB1 and EGFR TKIs, then added ki67 (a marker for proteins) and EdU (a marker for DNA). By using Zeiss Microscopy Software to quantify the cell images, this project gave insight into the correlation between the expression of KDM5A and E2F within the RB1 pathway.