FTO Inhibits 3T3-L1 Preadipocyte Differentiation by Regulation the m6A Modification of GPX4
10.13865/j.cnki.cjbmb.2025.09.1112
- VernacularTitle:脂肪和肥胖相关基因调节谷胱甘肽过氧化物酶4 m6 A修饰抑制3T3-L1前脂肪细胞分化
- Author:
Lin-Yuan HUANG
1
;
Jing GAO
;
Yi-Jin SUN
;
Hou-Jing JI
Author Information
1. 江苏卫生健康职业学院,医学基础部,生物化学教研室,南京 211800
- Publication Type:Journal Article
- Keywords:
fat mass and obesity associated gene(FTO);
m6A modification;
glutathione peroxidase 4(GPX4);
mitotic clonal expansion;
adipogenesis
- From:
Chinese Journal of Biochemistry and Molecular Biology
2025;41(11):1687-1699
- CountryChina
- Language:Chinese
-
Abstract:
The fat mass and obesity associated gene(FTO),a crucial RNA N6-methyladenosine(m6 A)demethylase,has been reported to influence the expression of glutathione peroxidase 4(GPX4)by modu-lating m6A modifications.GPX4 is a key molecule inhibiting ferroptosis,and the activation of ferroptosis signaling has been demonstrated to significantly reduce lipid accumulation in both mouse primary adipo-cytes and high fat diet fed mice.However,the specific m6A modification sites within the Gpx4 mRNA re-main undefined,and the regulatory role of Gpx4 during mouse adipocyte differentiation is also unclear.Through bioinformatic analysis combined with validation by methylated RNA immunoprecipitation sequen-cing(MeRIP)-qPCR and single-base elongation-and ligation-based qPCR amplification method(SE-LECT)assays,a key m6A modification site in Gpx4 mRNA was identified at 303 bp downstream from its transcription start site.CRISPR-Cas9-mediated knockdown of Gpx4 in 3T3-L1 cells,followed by adipo-genic induction,revealed that Gpx4 knockdown significantly reduced intracellular lipid droplet accumula-tion as assessed by Oil Red O staining(P＜0.001).RT-PCR and Western blotting analyses further dem-onstrated significantly decreased expression of key adipogenic differentiation genes(C/ebpα,Pparγ,Lpl,Fabp4)(P＜0.001).To investigate the temporal specificity of Gpx4 regulation,the GPX4 inhibitor RSL3(100 nmol/L)was administered during different stages of adipogenic differentiation.Results showed that RSL3 treatment specifically during the mitotic clonal expansion phase significantly suppressed the expression of adipogenic genes(Fabp4,Pparγ,Adipoq)and impeded adipogenesis.In summary,this study not only identifies a key m6A modification within the mouse Gpx4 mRNA but,more important-ly,reveals that GPX4 plays a critical regulatory role in 3T3-L1 adipocyte differentiation.These findings establish a link between the FTO-m6A-GPX4-ferroptosis regulatory axis and adipocyte differentiation,providing novel theoretical insights into the pathological mechanisms of obesity and identifying potential therapeutic targets.
