MiR-1a-3p Inhibits Apoptosis in Fluoride-exposed LS8 Cells by Targeting Map3k1

Abstract

Dental fluorosis is a common chemical disease. It is currently unclear how fluorosis occurs at the molecular level. We used miRNA-seq to look at the differences between miRNAs in the cell line of ameloblasts LS8 that had been treated with 3.2 mmol/L NaF. We also performed gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. miR-1a-3p levels were significantly lower in mouse LS8 cells treated with 3.2 mmol/L NaF, and miR-1a-3p-targeted genes were significantly enriched in the MAPK pathway. LS8 cells were divided into four groups: control, NaF, NaF+miR-1a-3p mimics, and NaF+miR-1a-3p mimics normal control groups. Cellular morphology was observed by an inverted microscope, and the proliferation activity of LS8 cells was assessed by Cell Counting Kit-8 (CCK-8). Using the real-time quantitative polymerase chain reaction (RT-qPCR), transcription levels of miR-1a-3p and Map3k1 were detected. The expressions of Bax, Bcl-2, Map3k1, p38MAPK, ERK1/2, p-p38MAPK, and p-ERK1/2 were measured by Western blot. After bioinformatics analysis, we used a luciferase reporter assay (LRA) to validate the target of miR-1a-3p, showing that miR-1a-3p could inhibit apoptosis while increasing proliferation in fluoride-exposed LS8 cells. Generally, miR-1a-3p might directly inhibit Map3k1, reduce MAPK signal pathway activation, and promote phosphorylation. Thus, our findings revealed that the interaction of miR-1a-3p with its target gene Map3k1 and MAPK signal pathway might decrease the apoptosis of LS8 cells treated with 3.2 mmol/L NaF.

Exploration of altered miRNA expression and function in MSC-derived extracellular vesicles in response to hydatid antigen stimulation

“…This may be due to species variability or different cell type responses. Other miRNAs, such as miR-124, miR-206-3p, miR-1a-3p, miR-134, miR-23a-3p, miR-199, miR-486-5p, miR-210-3p, and miR-185, showed differential expression after antigenic stimulation and are known to be involved in immune regulation, inflammation, osteogenic differentiation, angiogenesis, and apoptosis ( Li et al, 2016 ; Huang and Zhu, 2018 ; Zhao et al, 2018 , 2023 ; Chen et al, 2019 , 2023 ; Chen J. X. et al, 2020 ; Ghafouri-Fard et al, 2021 ; Yang et al, 2021 ; Zhang Z. et al, 2021 ; Wang et al, 2022 ; Castaño et al, 2023 ; Dong et al, 2023 ; Li and Li, 2023 ; Liu et al, 2023 ; Ma et al, 2023 ; Su et al, 2023 ).…”