Long non-coding RNA LINC00616 promotes ferroptosis of periodontal ligament stem cells via the microRNA-370 / transferrin receptor axis
Hongwei Wang, Xiaotong Qiao, Chao Zhang, Jingyi Hou, Suqing Qi
Abstract
This study was designed to explore the role of lncRNA LINC00616 in the regulation of periodontitis. Cellular functions were measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assays. The content of reactive oxygen species, Fe2+, glutathione, and malondialdehyde were measured to determine ferroptosis in Porphyromonas gingivalis lipopolysaccharide (LPS-PG) treated periodontal ligament stem cells (PDLSCs), as well as expression of glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11, and acyl-CoA synthetase long-chain family member 4 proteins mRNA and miRNA levels were measured by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Western blot analysis was performed to assess protein expression. Targeting relationships were predicted using StarBase and TargetScan and verified by a dual luciferase reporter assay. The lncRNA LINC00616 was upregulated in periodontitis ligament tissues of patients with periodontitis and in PDLSCs treated with LPS-PG. Inhibition of LINC00616 promoted cell viability and suppressed ferroptosis of PDLSCs. miR-370 was verified to be a target of LINC00616, and suppressed miR-370 reversed the effects of LINC00616 knockdown on cell viability and ferroptosis in PDLSCs. Additionally, miR-370 targeting the transferrin receptor protein and upregulated transferrin receptor (TFRC) abolished the effects of overexpressed miR-370 on cell viability and ferroptosis of PDLSCs. LINC00616 acted as a competitive endogenous RNA (ceRNA) to promote ferroptosis of PDLSCs via the miR-370/TFRC axis. Therefore, LINC00616 knockdown may be a promising therapeutic strategy for periodontitis.