[Lipopolysaccharide induces inflammation and up-regulates the expression of methyltransferase-like 3 (METTL3) and METTL14 in human and mouse intestinal epithelial cells].

ABSTRACT

Objective To investigate the changes of N6-methyladenosine (m6A) modification in the inflammatory status of HIEC-6 human intestinal epithelial cells and MODE-K mouse intestinal epithelial cells. Methods HIEC-6 cells and MODE-K cells were induced by different concentrations of lipopolysaccharide (LPS), interleukin-1ß (IL-1ß), IL-6 and tumor necrosis factor alpha (TNF-α) for 10 hours or the same concentration of LPS, IL-1ß, IL-6 and TNF-α for 0, 3, 6, 12, 24 hours, respectively. The mRNA expression levels of pro-inflammatory cytokines IL-1ß, IL-6 and TNF-α were detected by real-time quantitative PCR. The mRNA and protein expression levels of m6A modification-related molecules methyltransferase-like 3 (METTL3), METTL14, METTL16, Wilm's tumor 1-associated protein (WTAP), alkylation repair homolog protein 5 (ALKBH5), fat-mass and obesity-associated protein (FTO), YTH domain-containing 1 (YTHDC1), YTHDC2 were detected through real-time quantitative PCR and Western blot, respectively. Results The mRNA expression levels of IL-1ß, IL-6 and TNF-α were increased and the mRNA and protein expression levels of METTL3 and METTL14 were simultaneously up-regulated in time-dependent and concentration-dependent LPS-induced model in HIEC-6 cells and MODE-K cells. Conclusion LPS can induce inflammation and up-regulate the expression of METTL3 and METTL14 in intestinal epithelial cells.