Fluoride resistance capacity in mammalian cells involves global gene expression changes associate with ferroptosis

Objective: The purpose of this study was to understand mouse osteoblast Ferroptosis under high fluoride environment by stimulating fluoride levels to corresponding levels. In order to define the underlying mechanism of fluoride resistance in mammals and provide a theoretical basis for fluorosis treatment, high-throughput Sequencing was applied to map the genetic changes of fluoride-resistant mouse osteoblasts and analyze the role of ferroptosis-related genes.

Methods: Cell Counting Kit-8, Reactive Oxygen Species Assay Kit and C11 BODIPY 581/591 were used to monitor proliferation and Ferroptosis of mouse osteoblasts MC3T3-E1 under high fluoride environment. Fluoride-tolerant MC3T3-E1 cells were developed by gradient fluoride exposure. The differentially expressed genes of fluorine-resistant MC3T3-E1 cells were identified by high-throughput Sequencing.

Results: MC3T3-E1 cells cultured in medium containing 20, 30, 60, 90 ppm F^- exhibited decreased viability and increased Reactive Oxygen Species and lipid peroxidation levels in correlation with F^- concentrations. High-throughput RNA Sequencing identified 2702 differentially expressed genes (DEGs) showed more than 2-fold difference in 30 ppm FR MC3T3-E1 cells, of which 17 DEGs were associated with Ferroptosis.

Conclusion: High fluoride environment affected the content of lipid peroxides in the body and increased the level of Ferroptosis, further, ferroptosis-related genes played specific roles in the fluoride resistance of mouse osteoblasts.

Ferroptosis; Fluoride resistance; Fluorosis; Skeletal fluorosis.