METTL3/m6A/miRNA-873-5p Attenuated Oxidative Stress and Apoptosis in Colistin-Induced Kidney Injury by Modulating Keap1/Nrf2 Pathway

Abstract

Nephrotoxicity of colistin is the major factor limiting its clinical application. However, the exact mechanism of colistin-induced nephrotoxicity is still elusive. N 6 -Methyladenosine (m 6 A) modification has been implicated in many biological processes, however, its role in colistin-induced nephrotoxicity needs to be elucidated. Mouse renal tubular epithelial cells (mRTECs) were treated with 200 μM colistin with or without METTL3 overexpression. Cells injury, m 6 A assay, oxidative stress and apoptosis were examined. Levels of m 6 A are decreased after colistin treatment in mRTECs. METTL3 is the major factor involved in abnormal m 6 A modification. METTL3 overexpression plays a protective role against colistin-induced oxidative stress and apoptosis. Moreover, METTL3 interacts with the microprocessor protein DGCR8 and positively modulates miR-873-5p mature process in an m 6 A-dependent manner. Further experiments show that miR-873-5p could regulate Keap1-Nrf2 pathway against colistin-induced oxidative stress and apoptosis. These studies revealed an important role of METTL3/m 6 A in colistin-induced nephrotoxicity and provide a new insight on m 6 A modification in drug induced toxicity.

Multiple Functions and Mechanisms Underlying the Role of METTL3 in Human Cancers

“…The lncRNA ARHGAP5-AS1, miR-600, miRNA let-7g, and miR-33a can influence human cancer progression by targeting METTL3 (35,36,52,53). Conversely, METTL3 can promote pri-miRNA processing by interacting with DGCR8 (2,20,54). Dysregulation of METTL3 in various cancers can also influence cancer cell EMT (23,49,50), apoptosis (36,47), and stem cell self-renewal (38,39), which have been identified to be important in cancer progression.…”

“…Han et al found that METTL3 positively modulates pri-miR221/222 processing in an m 6 A-dependent manner by interacting with DGCR8, resulting in a reduction in the PTEN level, which ultimately results in proliferation of bladder cancer (29). Wang et al explored the function and mechanism of METTL3 and m 6 A in colistin-induced kidney injury and found that METTL3 interacts with DGCR8 and positively modulates the processing of mature miR-873-5p in an m 6 A-dependent manner (54). Collectively, this evidence suggests that DGCR8 plays an important role in the m 6 A-dependent regulation of miRNA processing by METTL3.…”

RNA N6-methyladenosine: a promising molecular target in metabolic diseases

“…So far, m 6 A has been found not only in mRNAs, but also in a variety of non-coding RNAs including rRNA, tRNA, snRNA, miRNA, and lncRNA [14,15]. For example, m 6 A methyltransferase-like 3 (METTL3) interacts with the microprocessor protein DGCR8 and modulates miR-873-5p mature process positively [16]. The expression of m 6 A demethylase fat mass and obesity-associated protein (FTO) can influence the steady state level of various miRNAs, including increased expression of hsa-miR-6505-5p, hsa-miR-651-5p and hsa-miR-493-5p, and decreased expression of hsa-miR-7-5p, hsa-miR-92a-1-5p and hsa-miR-6769a-3p [15].…”

7-Hydroxycoumarin Attenuates Colistin-Induced Kidney Injury in Mice Through the Decreased Level of Histone Deacetylase 1 and the Activation of Nrf2 Signaling Pathway

“…Western blotting was carried out as previously described (Wang et al, 2019). Nuclear proteins were extracted using a Nuclear Protein Extraction Kit (Solarbio, Beijing, China) and total protein were prepared in radio-immunoprecipitation assay (RIPA) lysis buffer containing 0.5 mM PMSF (Beyotime, Haimen, China), and centrifuged at 12,000 × g for 15 min at 4°C.…”

“…The procedure was carried out as described previously (Wang et al, 2019). Briefly, total RNA was extracted using Trizol reagent (Life Technologies, Grand Island, NY, USA) according to the manufacturer's instructions.…”