N6‐methyladenosine modification contributes to arecoline‐mediated oral submucosal fibrosis

Abstract

Background: Oral submucosal fibrosis (OSF) is a precancerous condition that closely related to the habit of chewing betel nut. The OSF patients of 3%-19% may develop cancer, and this probability is increasing year by year. Epigenetics modifications have been reported as part of the pathogenesis of OSF. However, in OSF field, the role and mechanism of arecoline-induced activation of transforming growth factor β (TGF-β) signaling on N6-methyladenosine (m6A) modification remain unclear. In this study, we investigated the effect and mechanism of arecoline on m6A modification.Methods: MeRIP-Seq and RNA-seq were performed in arecoline-stimulated cells.Quantitative polymerase chain reaction and western blot were performed to detect the expression of m6A writers and erasers. CCK-8 and flow cytometry analyses were performed to measure the cell viability and apoptosis.Results: m6A level was increased in OSF tissues compared to normal tissues; arecoline promoted the m6A methyltransferase Mettl3 and Mettl14 through TGF-β.MeRIP-seq and RNA-seq analyses found that MYC was the target gene of Mettl14.In addition, Mettl14 silence reversed the effects of arecoline on cell proliferation and apoptosis in Hacat cells.Conclusion: TGF-β-METTL14-m6A-MYC axis was crucially implicated in arecolinemediated OSF and may be an effective therapeutic strategy for OSF treatment.

DPSCs regulate epithelial-T cell interactions in oral submucous fibrosis

“…To investigate the possible interaction between Epi1.2 and T cells in the development of OSF, we established an Epi1.2 cell model of OSF in vitro using arecoline, a primary component implicated in the progression of OSF [ 35 , 36 ]. CCK8 assays and scratch tests were conducted to evaluate the impact of different concentrations of arecoline solution on the proliferation and migration of epithelial cells (HOK) over 48 h. The results showed that high concentrations of arecoline (60 µg/mL, 80 µg/mL) stimulated the proliferation of epithelial cells within the first 24 h. The proliferation ability of HOK cells could be decreased by a high concentration arecoline solution, as compared to the low concentration (20 µg/mL, 40 µg/mL) solution after 48 h (Fig.…”

Transcriptome-wide m6A methylome analysis uncovered the changes of m6A modification in oral pre-malignant cells compared with normal oral epithelial cells

“…However, the status of m 6 A modification and its relationship with m 6 A regulator expression have not been studied. Although the overall m 6 A modification was enhanced in precancerous oral submucosal fibrosis, the target genes and corresponding regulatory factors were unclear ( 29 ). In this study, we used a well-established precancerous cell line ( 30 ) as a model to investigate the potential contribution of m 6 A modification to the initiation of cell transformation.…”

“…Oral submucosal fibrosis (OSF) is a precancerous condition. Compared with normal tissues, the overall m 6 A level in OSF tissues was increased, suggesting that m 6 A modification contributes to OSF ( 29 ). However, to our knowledge, the change of m 6 A profile and the expression of m 6 A regulators in precancerous cells have not been well studied.…”

Transcription Factor TFAP2B Exerts Neuroprotective Effects Targeting BNIP3-Mediated Mitophagy in Ischemia/Reperfusion Injury

“…The relative gene expressions were calculated in accordance with the 2 ΔΔCt method. The methylated m6A RNA immunoprecipitation (me-RIP) assay was performed as described previously [24]. Relevant primer information was listed in Table 1.…”