O-GlcNAcylation drives calcium signaling toward osteoblast differentiation: A bioinformatics-oriented study

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3 Citations (Scopus)


This study aimed to reveal the possible mechanisms by which O-linked-N-acetylglucosaminylation (O-GlcNAcylation) regulates osteoblast differentiation using a series of bioinformatics-oriented experiments. To examine the influence of O-GlcNAcylation levels on osteoblast differentiation, osteoblastic MC3T3-E1 cells were treated with O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) inhibitors. Correlations between the levels of O-GlcNAcylation and the expression of osteogenic markers as well as OGT were evaluated by qPCR and western blotting. The O-GlcNAcylated proteins assumed to correlate with Runx2 expression were retrieved from several public databases and used for further bioinformatics analysis. Following the findings of the bioinformatics analysis, intracellular calcium ([Ca2+]i) was monitored in the cells treated with OGT and OGA inhibitors using a confocal laser-scanning microscope (CLS). The interaction effect between O-GlcNAcylation and [Ca2+]i on osteogenic marker expression was determined using stable OGT knockdown MC3T3-E1 cells. O-GlcNAcylation was positively associated with osteoblast differentiation. The time-course profile of global O-GlcNAcylated proteins showed a distinctive pattern with different molecular weights during osteoblast differentiation. The expression pattern of several O-GlcNAcylated proteins was significantly similar to that of Runx2 expression. Bioinformatic analysis of the retrieved Runx2-related-O-GlcNAcylated-proteins revealed the importance of [Ca2+]i. CLS showed that alteration of O-GlcNAcylation rapidly changed [Ca2+]i in MC3T3-E1 cells. O-GlcNAcylation and [Ca2+]i showed an interaction effect on the expression of osteogenic markers. OGT knockdown disrupted the [Ca2+]i-induced expression changes of osteogenic markers. O-GlcNAcylation interacts with [Ca2+]i and elicits osteoblast differentiation by regulating the expression of osteogenic markers.

Original languageEnglish
Pages (from-to)992-1015
Number of pages24
Issue number6
Publication statusPublished - Nov 1 2021


  • O-linked-N-acetylglucosaminylation
  • bioinformatics
  • calcium
  • literature mining
  • osteoblast differentiation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Clinical Biochemistry


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