Augmentation of Transforming Growth Factor-β-induced Epithelial to Mesenchymal Transition by Interleukin-1β: Role of DNA Methylation
Location
Philadelphia, PA
Start Date
30-4-2025 1:00 PM
End Date
30-4-2025 4:00 PM
Description
Lung cancer is a leading cause of cancer-related deaths worldwide. A better understanding of
mechanisms related to tumor initiation, progression and metastasis might help develop 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 (IL-1) enhances transforming growth factor beta (TGF-β) induced EMT in A549 lung adenocarcinoma cells, and whether regulation of DNA methylation is involved in this enhancement. Cells in 4 treatment groups (IL-1 alone, TGF-β alone, IL-1 and TGF-β cotreatment 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 andDNMT3a mRNA expression between TGF-β alone and TGF-β-IL1 cotreatment. However, IL-1 significantly decreased TET-1 mRNA expression as compared to untreated controls, and inhibited TGF-β induction of TET-1 at 24 and 48 hours. Future experiments will examine changes in global levels of 5mC and 5hmC, as well as DNMT and TET enzyme activity. We hypothesize that decreased TET activity facilitates early steps in EMT transition initiated by TGF-β.
Embargo Period
5-20-2025
Augmentation of Transforming Growth Factor-β-induced Epithelial to Mesenchymal Transition by Interleukin-1β: Role of DNA Methylation
Philadelphia, PA
Lung cancer is a leading cause of cancer-related deaths worldwide. A better understanding of
mechanisms related to tumor initiation, progression and metastasis might help develop 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 (IL-1) enhances transforming growth factor beta (TGF-β) induced EMT in A549 lung adenocarcinoma cells, and whether regulation of DNA methylation is involved in this enhancement. Cells in 4 treatment groups (IL-1 alone, TGF-β alone, IL-1 and TGF-β cotreatment 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 andDNMT3a mRNA expression between TGF-β alone and TGF-β-IL1 cotreatment. However, IL-1 significantly decreased TET-1 mRNA expression as compared to untreated controls, and inhibited TGF-β induction of TET-1 at 24 and 48 hours. Future experiments will examine changes in global levels of 5mC and 5hmC, as well as DNMT and TET enzyme activity. We hypothesize that decreased TET activity facilitates early steps in EMT transition initiated by TGF-β.