The prognosis for diffuse gliomas is very poor and the mechanism underlying their malignant progression remains unclear. Here, we aimed to elucidate the role and mechanism of the RNA N6,2′-O-dimethyladenosine (m6A) reader, YTH N6-methyladenosine RNA binding protein 2 (YTHDF2), in regulating the malignant progression of gliomas. YTHDF2 mRNA levels and functions were assessed using several independent datasets. Western blotting, quantitative polymerase chain reaction, and immunohistochemistry were used to evaluate the expression levels of YTHDF2 and other molecules in human and mouse tumor tissues and cells. Knockdown and overexpression were used to evaluate the effects of YTHDF2, methyltransferase-like 3 (METTL3), and UBX domain protein 1 (UBXN1) on glioma malignancy in cell and orthotopic xenograft models. RNA immunoprecipitation (RIP), methylated RIP, and RNA stability experiments were performed to study the mechanisms underlying the oncogenic role of YTHDF2. YTHDF2 expression was positively associated with a higher malignant grade and molecular subtype of glioma and poorer prognosis. YTHDF2 promoted the malignant progression of gliomas in both in vitro and in vivo models. Mechanistically, YTHDF2 accelerated UBXN1 mRNA degradation via METTL3-mediated m6A, which, in turn, promoted NF-κB activation. We further revealed that UBXN1 overexpression attenuated the oncogenic effect of YTHDF2 overexpression and was associated with better survival in patients with elevated YTHDF2 expression. Our findings confirmed that YTHDF2 promotes the malignant progression of gliomas and revealed important insight into the upstream regulatory mechanism of NF-κB activation via UBXN1 with a primary focus on m6A modification.

中文翻译：

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YTHDF2通过识别METTL3介导的m6A修饰来激活NF-κB并促进胶质瘤的恶性进展，从而促进UBXN1 mRNA衰变

弥漫性胶质瘤的预后很差，其恶性进展的机制尚不清楚。在这里，我们旨在阐明 RNA N6,2'-O-二甲基腺苷 (m6A) 阅读器 YTH N6-甲基腺苷 RNA 结合蛋白 2 (YTHDF2) 在调节胶质瘤恶性进展中的作用和机制。使用几个独立的数据集评估 YTHDF2 mRNA 水平和功能。使用蛋白质印迹、定量聚合酶链反应和免疫组织化学来评估 YTHDF2 和其他分子在人和小鼠肿瘤组织和细胞中的表达水平。敲低和过表达用于评估 YTHDF2、甲基转移酶样 3 (METTL3) 和 UBX 域蛋白 1 (UBXN1) 在细胞和原位异种移植模型中对胶质瘤恶性肿瘤的影响。RNA免疫沉淀（RIP），进行甲基化 RIP 和 RNA 稳定性实验以研究 YTHDF2 致癌作用的潜在机制。YTHDF2表达与胶质瘤较高的恶性分级和分子亚型以及较差的预后呈正相关。YTHDF2在体外和体内模型中促进了胶质瘤的恶性进展。从机制上讲，YTHDF2 通过 METTL3 介导的 m6A 加速 UBXN1 mRNA 降解，进而促进 NF-κB 活化。我们进一步揭示了 UBXN1 过表达减弱了 YTHDF2 过表达的致癌作用，并且与 YTHDF2 表达升高的患者更好的生存相关。