Dataset Information

Long non-coding RNA LINC00616 promotes ferroptosis of periodontal ligament stem cells via the microRNA-370 / transferrin receptor axis.

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.

SUBMITTER: Wang H

PROVIDER: S-EPMC9276003 | biostudies-literature | 2022 May

REPOSITORIES: biostudies-literature

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Long non-coding RNA LINC00616 promotes ferroptosis of periodontal ligament stem cells via the microRNA-370 / transferrin receptor axis.

Wang Hongwei H Qiao Xiaotong X Zhang Chao C Hou Jingyi J Qi Suqing S

Bioengineered 20220501 5

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, Fe<sup>2+</sup>, glutathione, and malondialdehyde were measured to determine ferroptosis in <i>Porphyromonas gingivalis</i> lipopolysaccharide (LPS-PG) treated peri ...[more]

PMID: 35611986

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Project description:BackgroundLong non-coding RNAs (lncRNAs) have been widely known to have an appreciable effect in physiology and pathology. In tooth regeneration, periodontal ligament stem cells (PDLSCs) are regarded as a key effector, whereas, how lncRNA acts in the osteogenic differentiation of PDLSCs have not been completely understood. This study aims to find out the relationship between lncRNA DANCR and the proliferation and osteogenic differentiation of PDLSCs.MethodsMicroarray was used to observe the different expression of lncRNAs in differentiated and undifferentiated PDLSCs. And then osteogenic-related lncRNA, DANCR was screened out. Its effects on proliferation and osteogenic differentiation was explored by constructing an overexpression and inhibition model. qRT-PCR was used to detect the mRNA expression of osteogenesis related genes. MTT assay was performed to assess the effects of DANCR on cell growth curve. To quantify the effects of DANCR on osteogenic differentiation of PDLSCs, ALP staining and alizarin red was performed in basic culture medium and osteogenic medium. Data were statistically processed.ResultsCompared with the undifferentiated PDLSCs, the alizarin red staining level was higher in differentiated PDLSCs. And the expressions of osteogenic differentiation marker genes Runt-related transcription factor 2 (Runx2), osteocalcin (OCN) and bone morphogenetic protein (BMP-2) were significantly increased in the differentiated PDLSCs. Furthermore, we noticed that comparing with control groups, the expression of lncRNA DANCR decreases markedly in osteogenically induced PDLSCs. DANCR promoted proliferation of PDLSCs, as evidenced by cell viability. Further investigation has proven that the downregulation of DANCR shows in the calcium sediment forming, alkaline phosphatase (ALP) activation and some osteogenic-related gene markers' upregulation including Runx2, OCN and BMP-2, which finally results in the osteogenic differentiation of PDLSCs following the transfection and induction. Conversely, DANCR upregulation was shown to repress the osteogenic differentiation potential of PDLSCs.ConclusionsThe osteogenic differentiation of PDLSCs has proven to related to the down regulation of lncRNA DANCR. And this paper throws light on the effects of DANCR in the process of PDLSCs' osteogenic differentiation.

Dataset Information

Long non-coding RNA LINC00616 promotes ferroptosis of periodontal ligament stem cells via the microRNA-370 / transferrin receptor axis.

Publications

Long non-coding RNA LINC00616 promotes ferroptosis of periodontal ligament stem cells via the microRNA-370 / transferrin receptor axis.

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