Project Overview
The role of KLF4 in genomic stability
Department(s)
Biology
Abstract
PROJECT 1: Krüppel-like factor four (Klf4) is a zinc-finger transcription factor that is involved in cell proliferation, differentiation, autophagy, and genomic stability. It has been implicated as a hallmark of colorectal cancers, one of the most common cancers worldwide. Our recent findings indicated that in the absence of Klf4, damage to mitochondria may facilitate a shift toward glycolytic metabolism, an indicative of the Warburg effect. However, the role of Klf4 in lipid metabolism as a method of maintaining genomic integrity remains to be elucidated. This study seeks to address the role of KLF4 in lipid metabolism in both mouse embryonic fibroblasts (MEFs) and human colorectal cancer cells (RKOs). To investigate our research question, we will treat cells with a glycolytic inhibitor and 2-deoxy-D-glucose and lactate dehydrogenase A inhibitor and perform a protein assay (western blot) for lipid metabolism genes involved in fatty acid oxidation. In addition, we will do an ATP assay to determine energy efficiency in cells lacking klf4 as compared with wild-type. Taken together, our findings will provide us a rationale for the impaired metabolism in cells-null for klf4, and Klf4 regulates various metabolic enzymes to maintain metabolism homeostasis.
PROJECT 2: The dysregulation of normal cellular processes, such as the evasion of programmed cell death, often characterizes cancer. With this, ferroptosis is a newly discovered form of iron-dependent, programmed cell death characterized by excess reactive oxygen species and lipid peroxides within the cell. In recent studies, Krüppel-like factor 4 (KLF4), a zinc transcription factor, has been hypothesized to regulate ferroptosis-related genes. However, the mechanism behind which KLF4 functions in ferroptotic pathways remains unknown. In this study, we aim to determine if KLF4 regulates ferroptosis in mouse embryonic fibroblasts (MEFs) and RKO cells. To study KLF4’s potential role in the regulation of ferroptosis-related genes, focusing on important genes in the glutathione dependent pathway, xCT/SLC7A11 and GPX4, KLF4 wildtype and null MEFs were treated for 72 hours with erastin, a ferroptosis-inducing drug, and DMSO, as a control. After sample collection, western blot analysis will be conducted to determine if there will vary in the expression levels of ferroptosis-related genes between the KLF4 wildtype and null MEFs.
Student Qualifications
Must complete BIOLOGY 182, Knowledge of cell culture and western blot preferable
Project Length
10 weeks
APPLY