DOI
https://doi.org/10.25772/F2P0-DJ38
Defense Date
2024
Document Type
Thesis
Degree Name
Master of Science
Department
Human and Molecular Genetics
First Advisor
Dr. Devanand Sarkar
Second Advisor
Dr. Zheng Fu
Third Advisor
Dr. Lauren Cowart
Abstract
Hepatocellular carcinoma (HCC) is the most common primary liver cancer and a leading cause of cancer-related deaths worldwide. Despite advances in understanding its molecular pathogenesis, treatment options for HCC remain limited, highlighting the need for identifying novel therapeutic targets. This study investigates the role of SLC25A32, a mitochondrial transporter gene located on chromosome 8q22.3, in HCC progression. SLC25A32 is involved in the transport of tetrahydrofolate (THF) and flavin adenine dinucleotide (FAD) into mitochondria, playing a crucial role in one-carbon metabolism and redox balance. Using CRISPR/Cas9 technology, we generated potential SLC25A32 knockout clones in the HCC cell line Huh-7. Quantitative PCR analysis revealed downregulation of SLC25A32 expression in these clones compared to control cells. In parallel, we evaluated SLC25A32 overexpression using cell lines provided by our collaborator and observed increased gene expression levels and enhanced colony formation, suggesting a role in promoting cell proliferation. To further elucidate the functional consequences of SLC25A32 modulation, we performed knockdown experiments in the Hep3B cell line and assessed changes in proliferation rates using colony formation assays. Additionally, we attempted to validate SLC25A32 protein expression levels through Western blotting but faced challenges due to the lack of reliable antibodies. Our findings support the hypothesis that SLC25A32 plays a crucial role in HCC progression, potentially through its involvement in regulating cellular metabolism and proliferation. However, the ongoing controversy surrounding its primary function as an FAD transporter or a folate transporter underscores the need for further investigation into the specific mechanisms underlying its effects in HCC. Future studies will focus on comprehensive functional assays, metabolomics analysis, in vivo tumorigenesis models, and the exploration of SLC25A32 as a potential therapeutic target or biomarker for HCC. Understanding the intricate role of SLC25A32 in HCC biology may contribute to the development of more effective and personalized treatment strategies for this aggressive malignancy.
Rights
© The Author
Is Part Of
VCU University Archives
Is Part Of
VCU Theses and Dissertations
Date of Submission
5-9-2024