Fumonisin B Alters Global M6A RNA Methylation and Epigenetically Regulates Keap1-Nrf2 Signaling in Human Hepatoma (HepG2) Cells

Overview

Journal Arch Toxicol

Specialty Toxicology

Date 2021 Jan 26

PMID 33496827

Citations 14

Authors

Thilona Arumugam

Terisha Ghazi

Anil A Chuturgoon

Affiliations

Soon will be listed here.

Abstract

FB is a common contaminant of cereal grains that affects human and animal health. It has become increasingly evident that epigenetic changes are implicated in FB toxicity. N6-methyladenosine (m6A), the most abundant post-transcriptional RNA modification, is influenced by fluctuations in redox status. Since oxidative stress is a characteristic of FB exposure, we determined if there is cross-talk between oxidative stress and m6A in FB-exposed HepG2 cells. Briefly, HepG2 cells were treated with FB (0, 5, 50, 100, 200 µM; 24 h) and ROS, LDH and m6A levels were quantified. qPCR was used to determine the expression of m6A modulators, Nrf2, Keap1 and miR-27b, while western blotting was used to quantify Keap1 and Nrf2 protein expression. Methylation status of Keap1 and Nrf2 promoters was assessed and RNA immunoprecipitation quantified m6A-Keap1 and m6A-Nrf2 levels. FB induced accumulation of intracellular ROS (p ≤ 0.001) and LDH leakage (p ≤ 0.001). Elevated m6A levels (p ≤ 0.05) were accompanied by an increase in m6A "writers" [METLL3 (p ≤ 0.01) and METLL14 (p ≤ 0.01)], and "readers" [YTHDF1 (p ≤ 0.01), YTHDF2 (p ≤ 0.01), YTHDF3 (p ≤ 0.001) and YTHDC2 (p ≤ 0.01)] and a decrease in m6A "erasers" [ALKBH5 (p ≤ 0.001) and FTO (p ≤ 0.001)]. Hypermethylation and hypomethylation occurred at Keap1 (p ≤ 0.001) and Nrf2 (p ≤ 0.001) promoters, respectively. MiR-27b was reduced (p ≤ 0.001); however, m6A-Keap1 (p ≤ 0.05) and m6A-Nrf2 (p ≤ 0.01) levels were upregulated. This resulted in the ultimate decrease in Keap1 (p ≤ 0.001) and increase in Nrf2 (p ≤ 0.001) expression. Our findings reveal that m6A RNA methylation can be modified by exposure to FB, and a cross-talk between m6A and redox regulators does occur.

Citing Articles

The interplay between RNA m6A modification and radiation biology of cancerous and non-cancerous tissues: a narrative review.

Cheng Y, Shang Y, Zhang S, Fan S Cancer Biol Med. 2025; 21(12.

PMID: 39831771 PMC: 11745087. DOI: 10.20892/j.issn.2095-3941.2024.0415.

Ferroptosis: a critical mechanism of N-methyladenosine modification involved in carcinogenesis and tumor progression.

Wei Q, Xue C, Li M, Wei J, Zheng L, Chen S Sci China Life Sci. 2024; 67(6):1119-1132.

PMID: 38811442 DOI: 10.1007/s11427-023-2474-4.

Revisiting Neuroblastoma: Nrf2, NF-κB and Phox2B as a Promising Network in Neuroblastoma.

Peggion S, Najem S, Kolman J, Reinshagen K, Pagerols Raluy L Curr Issues Mol Biol. 2024; 46(4):3193-3208.

PMID: 38666930 PMC: 11048850. DOI: 10.3390/cimb46040200.

Characterization of the mA regulators' landscape highlights the clinical significance of acute myocardial infarction.

Chao P, Zhang X, Zhang L, Wang Y, Wusiman M, Aimaijiang G Front Immunol. 2024; 15:1308978.

PMID: 38571952 PMC: 10987706. DOI: 10.3389/fimmu.2024.1308978.

Multilevel regulation of N-methyladenosine RNA modifications: Implications in tumorigenesis and therapeutic opportunities.

Feng L, Du R, Chang B, Li M, Tian J, Wang S Genes Dis. 2023; 10(5):1969-1981.

PMID: 37492716 PMC: 10363589. DOI: 10.1016/j.gendis.2022.08.018.

References

Arumugam T, Pillay Y, Ghazi T, Nagiah S, Abdul N, Chuturgoon A . Fumonisin B-induced oxidative stress triggers Nrf2-mediated antioxidant response in human hepatocellular carcinoma (HepG2) cells. Mycotoxin Res. 2018; 35(1):99-109. DOI: 10.1007/s12550-018-0335-0. View

Chuturgoon A, Phulukdaree A, Moodley D . Fumonisin B1 induces global DNA hypomethylation in HepG2 cells - An alternative mechanism of action. Toxicology. 2013; 315:65-9. DOI: 10.1016/j.tox.2013.11.004. View

Chuturgoon A, Phulukdaree A, Moodley D . Fumonisin B₁ modulates expression of human cytochrome P450 1b1 in human hepatoma (Hepg2) cells by repressing Mir-27b. Toxicol Lett. 2014; 227(1):50-5. DOI: 10.1016/j.toxlet.2014.02.026. View

Demirel G, Alpertunga B, Ozden S . Role of fumonisin B1 on DNA methylation changes in rat kidney and liver cells. Pharm Biol. 2015; 53(9):1302-10. DOI: 10.3109/13880209.2014.976714. View

Desrosiers R, Friderici K, Rottman F . Identification of methylated nucleosides in messenger RNA from Novikoff hepatoma cells. Proc Natl Acad Sci U S A. 1974; 71(10):3971-5. PMC: 434308. DOI: 10.1073/pnas.71.10.3971. View

Dominissini D, Moshitch-Moshkovitz S, Schwartz S, Salmon-Divon M, Ungar L, Osenberg S . Topology of the human and mouse m6A RNA methylomes revealed by m6A-seq. Nature. 2012; 485(7397):201-6. DOI: 10.1038/nature11112. View

Eades G, Yang M, Yao Y, Zhang Y, Zhou Q . miR-200a regulates Nrf2 activation by targeting Keap1 mRNA in breast cancer cells. J Biol Chem. 2011; 286(47):40725-33. PMC: 3220489. DOI: 10.1074/jbc.M111.275495. View

Fabrizio F, Sparaneo A, Trombetta D, Muscarella L . Epigenetic versus Genetic Deregulation of the KEAP1/NRF2 Axis in Solid Tumors: Focus on Methylation and Noncoding RNAs. Oxid Med Cell Longev. 2018; 2018:2492063. PMC: 5872633. DOI: 10.1155/2018/2492063. View

Gelderblom W, Abel S, Smuts C, Marnewick J, Marasas W, Lemmer E . Fumonisin-induced hepatocarcinogenesis: mechanisms related to cancer initiation and promotion. Environ Health Perspect. 2001; 109 Suppl 2:291-300. PMC: 1240678. DOI: 10.1289/ehp.01109s2291. View

10.

Ghazi T, Nagiah S, Chuturgoon A . Fusaric acid decreases p53 expression by altering promoter methylation and m6A RNA methylation in human hepatocellular carcinoma (HepG2) cells. Epigenetics. 2020; 16(1):79-91. PMC: 7889137. DOI: 10.1080/15592294.2020.1788324. View

11.

Ghazi T, Nagiah S, Dhani S, Chuturgoon A . Fusaric acid-induced epigenetic modulation of hepatic H3K9me3 triggers apoptosis and . Epigenomics. 2020; 12(11):955-972. DOI: 10.2217/epi-2019-0284. View

12.

Han M, Liu Z, Xu Y, Liu X, Wang D, Li F . Abnormality of m6A mRNA Methylation Is Involved in Alzheimer's Disease. Front Neurosci. 2020; 14:98. PMC: 7058666. DOI: 10.3389/fnins.2020.00098. View

13.

Deng J, Zhang H, Li C, Huang H, Liu S, Yang H . 46,XX Testicular Disorders of Sex Development With Gene Mutation: First Case Report Identified Prenatally by Integrated Analyses in China. Front Genet. 2020; 10:1350. PMC: 7045042. DOI: 10.3389/fgene.2019.01350. View

14.

Karkowski M . [Stevens-Johnson syndrome with a fulminant course]. Wiad Lek. 1988; 41(5):317-9. View

15.

Kang K, Piao M, Kim K, Kang H, Chang W, Park I . Epigenetic modification of Nrf2 in 5-fluorouracil-resistant colon cancer cells: involvement of TET-dependent DNA demethylation. Cell Death Dis. 2014; 5:e1183. PMC: 4001304. DOI: 10.1038/cddis.2014.149. View

16.

Kouadio J, Dano S, Moukha S, Mobio T, Creppy E . Effects of combinations of Fusarium mycotoxins on the inhibition of macromolecular synthesis, malondialdehyde levels, DNA methylation and fragmentation, and viability in Caco-2 cells. Toxicon. 2006; 49(3):306-17. DOI: 10.1016/j.toxicon.2006.09.029. View

17.

Liu X, Fan L, Yin S, Chen H, Hu H . Molecular mechanisms of fumonisin B1-induced toxicities and its applications in the mechanism-based interventions. Toxicon. 2019; 167:1-5. DOI: 10.1016/j.toxicon.2019.06.009. View

18.

Livak K, Schmittgen T . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2002; 25(4):402-8. DOI: 10.1006/meth.2001.1262. View

19.

Meyer K, Saletore Y, Zumbo P, Elemento O, Mason C, Jaffrey S . Comprehensive analysis of mRNA methylation reveals enrichment in 3' UTRs and near stop codons. Cell. 2012; 149(7):1635-46. PMC: 3383396. DOI: 10.1016/j.cell.2012.05.003. View

20.

Mobio T, Anane R, Baudrimont I, Carratu M, Shier T, Dano S . Epigenetic properties of fumonisin B(1): cell cycle arrest and DNA base modification in C6 glioma cells. Toxicol Appl Pharmacol. 2000; 164(1):91-6. DOI: 10.1006/taap.2000.8893. View