Augmentation of Transforming Growth Factor-β-induced Epithelial to Mesenchymal Transition by Interleukin-1β: Role of DNA Methylation

Date of Award

6-2025

Degree Type

Thesis

Degree Name

Master of Science in Biomedical Sciences

First Advisor

Ruth Borghaei

Second Advisor

Dianzheng Zhang

Third Advisor

Brian DeHaven

Abstract

Lung cancer is a leading cause of cancer-related deaths worldwide. A better understanding of mechanisms related to tumor initiation, progression and metastasis could lead to better prevention and treatment regimens. Epithelial to mesenchymal transition (EMT) is related to metastasis and the development of therapy resistance. The aim of our study is to determine how inflammatory cytokine Interleukin-1 beta (IL-1β) enhances transforming growth factor beta (TGF-β) induced EMT in A549 lung adenocarcinoma cells, and whether regulation of DNA methylating or demethylating enzymes are involved in this enhancement. Cells in four treatment groups (IL-1β alone, TGF-β alone, IL-1β and TGF-β co-treatment and untreated control) were analyzed over a 96-hour period. Total RNA and genomic DNA were isolated at 24, 48, 72 and 96 hours. RT-PCR was used to measure mRNA levels of DNA methylating and DNA demethylating enzymes as well as established markers of EMT progression. Results of RT-PCR showed no significant difference in DNMT1 and DNMT3a mRNA expression between TGF-β alone and TGF-β-IL-1β cotreatment. However, IL-1β significantly decreased TET-1 mRNA expression as compared to untreated controls, and inhibited TGF-β induction of TET-1 at 24 and 48 hours. Inhibition of TET activity augmented TGF- β induced morphological changes but had no significant effect on CDH1/Ecadherin mRNA expression at 24 or 48 hours. Inhibition of cyclooxygenase-2 had no effect on the ability of IL-1β to augment TGF-β induced EMT.

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