Fluoride-Mediated Gene Expression Regulated Epigenetically via H3ac Modification in Ameloblast-Lineage Cells
Abstract
Objectives: Previously we reported that fluoride-mediated histone acetyltransferase (HAT) activation contributed to p53 acetylation to promote fluoride toxicity in mouse ameloblast-like cells (LS8). However, the roles of fluoride-mediated HAT activation in histone acetylation and epigenetic regulation of fluoride-mediated gene expressions remain unidentified. This study demonstrates that fluoride-mediated gene expression is regulated epigenetically via modification of histone acetylation status (H3K27Ac) in transcription start sites (TSS) in LS8 cells.
Methods: LS8 cells were treated with or without 5 mM fluoride for 24 h and subjected to chromatin isolation followed by immunoprecipitation (ChIP) and DNA purification. ChIP assay was performed using antibodies specific for H3K27Ac followed by ChIP-sequencing. Genes were identified by differential H3K27Ac peaks with fluoride treatment within ± 1 kb from TSS. Identified genes were analyzed by quantitative real-time PCR (q-PCR) to evaluate mRNA expressions.
Results: The differential acetylation status of H3K27 was associated with mRNA expressions (Bax, p21, Mdm2, p53, Bad and Bcl2) that were altered by fluoride. Fluoride increased H3K27Ac peaks of Bax, p21 and Mdm2, while decreased H3K27Ac peaks in p53, Bad and Bcl2. qPCR results showed that fluoride increased mRNAs of Bax, p21 and Mdm2, while suppressed mRNAs of p53, Bad and Bcl2. The H3K27Ac status (increase or decrease by fluoride) in these genes was concordant with mRNA expressions.
Conclusions: Gene expression altered by fluoride was epigenetically regulated via H3K27Ac inLS8 cells. Our results warrant further investigation to elucidate epigenetic regulation in fluoride toxicity to develop a potential novel strategy targeting histone modification.
SUPPORT FUNDING/GRANT NUMBER: NIH grant 1t34gm145509-01a1
Faculty Sponsors
Dr. Natsumi Fujiwara (Tokushima University Graduate School of Biomedical Sciences), Dr. Eric T. Everett (University of North Carolina at Chapel Hill), Dr. John D. Bartlett (The Ohio State University), Dr. Maiko Suzuki (Nova Southeastern University)
Project Type
Event
Location
Alvin Sherman Library
Start Date
4-3-2024 12:30 PM
End Date
4-4-2024 1:30 PM
Fluoride-Mediated Gene Expression Regulated Epigenetically via H3ac Modification in Ameloblast-Lineage Cells
Alvin Sherman Library
Objectives: Previously we reported that fluoride-mediated histone acetyltransferase (HAT) activation contributed to p53 acetylation to promote fluoride toxicity in mouse ameloblast-like cells (LS8). However, the roles of fluoride-mediated HAT activation in histone acetylation and epigenetic regulation of fluoride-mediated gene expressions remain unidentified. This study demonstrates that fluoride-mediated gene expression is regulated epigenetically via modification of histone acetylation status (H3K27Ac) in transcription start sites (TSS) in LS8 cells.
Methods: LS8 cells were treated with or without 5 mM fluoride for 24 h and subjected to chromatin isolation followed by immunoprecipitation (ChIP) and DNA purification. ChIP assay was performed using antibodies specific for H3K27Ac followed by ChIP-sequencing. Genes were identified by differential H3K27Ac peaks with fluoride treatment within ± 1 kb from TSS. Identified genes were analyzed by quantitative real-time PCR (q-PCR) to evaluate mRNA expressions.
Results: The differential acetylation status of H3K27 was associated with mRNA expressions (Bax, p21, Mdm2, p53, Bad and Bcl2) that were altered by fluoride. Fluoride increased H3K27Ac peaks of Bax, p21 and Mdm2, while decreased H3K27Ac peaks in p53, Bad and Bcl2. qPCR results showed that fluoride increased mRNAs of Bax, p21 and Mdm2, while suppressed mRNAs of p53, Bad and Bcl2. The H3K27Ac status (increase or decrease by fluoride) in these genes was concordant with mRNA expressions.
Conclusions: Gene expression altered by fluoride was epigenetically regulated via H3K27Ac inLS8 cells. Our results warrant further investigation to elucidate epigenetic regulation in fluoride toxicity to develop a potential novel strategy targeting histone modification.
SUPPORT FUNDING/GRANT NUMBER: NIH grant 1t34gm145509-01a1
