Background
Solute carrier family 7 member 11 (SLC7A11) is overexpressed in multiple human tumours and functions as a transporter importing cystine for glutathione biosynthesis. It promotes tumour development in part by suppressing ferroptosis, a newly identified form of cell death that plays a pivotal role in the suppression of tumorigenesis. However, the role and underlying mechanisms of SLC7A11‐mediated ferroptosis in hepatoblastoma (HB) remain largely unknown.
Methods
Reverse transcription quantitative real‐time PCR (RT‐qPCR) and western blotting were used to measure SLC7A11 levels. Cell proliferation, colony formation, lipid reactive oxygen species (ROS), MDA concentration, 4‐HNE, GSH/GSSG ratio and cell death assays as well as subcutaneous xenograft experiments were used to elucidate the effects of SLC7A11 in HB cell proliferation and ferroptosis. Furthermore, MeRIP‐qPCR, dual luciferase reporter, RNA pulldown, RNA immunoprecipitation (RIP) and RACE‐PAT assays were performed to elucidate the underlying mechanism through which SLC7A11 was regulated by the m6A modification in HB.
Results
SLC7A11 expression was highly upregulated in HB. SLC7A11 upregulation promoted HB cell proliferation in vitro and in vivo, inhibiting HB cell ferroptosis. Mechanistically, SLC7A11 mRNA exhibited abnormal METTL3‐mediated m6A modification, which enhanced its stability and expression. IGF2 mRNA‐binding protein 1 (IGF2BP1) was identified as the m6A reader of SLC7A11, enhancing SLC7A11 mRNA stability and expression by inhibiting SLC7A11 mRNA deadenylation in an m6A‐dependent manner. Moreover, IGF2BP1 was found to block BTG2/CCR4‐NOT complex recruitment via competitively binding to PABPC1, thereby suppressing SLC7A11 mRNA deadenylation.
Conclusions
Our findings demonstrated that the METTL3‐mediated SLC7A11 m6A modification enhances HB ferroptosis resistance. The METTL3/IGF2BP1/m6A modification promotes SLC7A11 mRNA stability and upregulates its expression by inhibiting the deadenylation process. Our study highlights a critical role of the m6A modification in SLC7A11‐mediated ferroptosis, providing a potential strategy for HB therapy through blockade of the m6A‐SLC7A11 axis.
Asphalt mastic plays an important role in asphalt mixtures for pavement engineering. Understanding the asphalt–filler interaction behavior is essential to improve the pavement performance of asphalt mastics. The objective of this paper was to evaluate the asphalt–filler interaction ability based on macro-rheological measurements and to investigate the asphalt–filler interaction mechanism associated with microstructural characteristics of asphalt mastics. First, the asphalt–filler interaction was characterized using macro-rheological features of asphalt mastics based on dynamic shear rheometer (DSR) tests. Second, the physico-chemical interaction between the asphalt and filler was qualitatively evaluated using a Fourier transform infrared (FTIR) spectrometer. Third, the asphalt–filler interaction behavior was investigated in terms of the micro-morphological properties of mineral fillers and asphalt mastics by conducting scanning electron microscopy (SEM) and atomic force microscope (AFM) tests. Finally, the grey relational analysis (GRA) was employed to identify the correlation between the properties of mineral fillers and the macro–micro performances of asphalt mastics. The results show that a higher content of alkaline mineral filler within the critical volume fraction range produced a greater interaction ability between the asphalt and filler. The asphalt–filler interaction is mainly a physical action since no obvious new adsorption peaks appeared in the FTIR spectrum. The micro-morphological characteristics of asphalt mastic mainly depended on the adsorption effect of mineral fillers on polar fractions and the dispersion effect of mineral fillers on wax crystals in the asphalt binder. Based on the results of the GRA, the acidity and content of mineral fillers exhibited a great influence on the micro-morphological and macro-rheological characteristics of asphalt mastics, and the specific surface area of the mineral filler exerted a significant influence on the asphalt–filler interaction ability. Furthermore, the K-B- δ index was more appropriate for evaluating the asphalt–filler interaction ability.
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