Department of Molecular Neuropathology, Department of Neuropathology, Beijing Neurosurgical Institute, Capital Medical University, No. 119 Nan Si Huan Xi Road, Fengtai District, Beijing, 100050, China.
Chinese Glioma Genome Atlas Network (CGGA), Beijing, China.
J Hematol Oncol. 2021 Jul 10;14(1):109. doi: 10.1186/s13045-021-01124-z.
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 (mA) 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 mA, 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 mA modification.
弥漫性神经胶质瘤的预后非常差，其恶性进展的机制仍不清楚。在这里，我们旨在阐明 RNA N6,2'-O-二甲基腺苷(mA)阅读器 YTH N6-甲基腺苷 RNA 结合蛋白 2(YTHDF2)在调节神经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 YTHDF2mRNA 水平和功能。Western blot、定量聚合酶链反应和免疫组织化学用于评估人及鼠肿瘤组织和细胞中 YTHDF2 和其他分子的表达水平。敲低和过表达用于评估 YTHDF2、甲基转移酶样 3(METTL3)和 UBX 结构域蛋白 1(UBXN1)对细胞和原位异种移植模型中神经胶质瘤恶性程度的影响。RNA 免疫沉淀(RIP)、甲基化 RIP 和 RNA 稳定性实验用于研究 YTHDF2 致癌作用的机制。
YTHDF2 表达与神经胶质瘤的恶性程度较高和分子亚型较高以及预后较差呈正相关。YTHDF2 在体外和体内模型中均促进神经胶质瘤的恶性进展。从机制上讲，YTHDF2 通过 METTL3 介导的 mA 加速 UBXN1mRNA 降解，从而促进 NF-κB 激活。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 高表达患者的生存改善有关。
我们的研究结果证实了 YTHDF2 促进神经胶质瘤的恶性进展，并揭示了 NF-κB 激活通过 UBXN1 的上游调节机制的重要见解，主要集中在 mA 修饰上。
