Global gene expression changes in osteoblasts from fluoride-tolerant mice

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. 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. High-throughput RNA sequencing identified 2702 differentially expressed genes (DEGs) showed more than 2-fold difference in 30ppm FR MC3T3-E1 cells, of which 17 DEGs were associated with ferroptosis. Wild-type MC3T3-E1 (WT MC3T3-E1) cells were continuously exposed to a gradient of 10 ppm F- to induced them survive in fluorinated medium. During cell adaptation, the cell proliferation was continuously monitored using Cell Counting Kit-8 (CCK-8) kit until a normal cell proliferation rate was confirmed. The above procedure was repeated for all other MC3T3-E1 clones to develop desired fluorine resistance. The differentially expressed genes of fluorine-resistant MC3T3-E1 cells were identified by high-throughput sequencing.