Yu Xiaofeng, Zhao He, Cao Zhiwei
Department of Otorhinolaryngology, Head and Neck Surgery, Shengjing Hospital of China Medical University, Shenyang, China.
Department of Otorhinolaryngology, Head and Neck Surgery, Shengjing Hospital of China Medical University, Shenyang, China -
Minerva Med. 2022 Apr;113(2):309-314. doi: 10.23736/S0026-4806.20.06653-7. Epub 2020 Jun 5.
This study aims to elucidate the role of METTL3 in aggravating the progression of NPC through mA modification on Snail and thus the stimulated epithelial-mesenchymal transition (EMT).
Differential expressions of METTL3 in 48 paired NPC tissues and paracancerous tissues were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Its level in NPC patients with different clinical stages and metastatic states was examined. Prognostic potential of METTL3 in NPC patients was assessed by Kaplan-Meier method. After knockdown of METTL3, expression changes of Snail and EMT-related genes, as well as invasive and migratory abilities in SUNE-1 cells were detected. The interaction between Snail with METTL3 and IGF2BP2 was verified by RIP (RNA-Binding Protein Immunoprecipitation) assay. At last, the roles of METTL3/Snail regulatory loop in influencing EMT and metastasis of NPC were clarified.
METTL3 was upregulated in NPC tissues than that of paracancerous ones. NPC patients with advanced stage or lymphatic metastasis expressed higher level of METTL3. Kaplan-Meier curves revealed that NPC patients expressing high level of METTL3 suffered worse prognosis. Knockdown of METTL3 downregulated protein levels of Snail and N-cadherin, while E-cadherin was upregulated in SUNE-1 cells. Meanwhile, knockdown of METTL3 inhibited invasive and migratory abilities in NPC cells. RIP assay confirmed the interaction between Snail and METTL3. Besides, knockdown of METTL3 decreased the enrichment abundance of Snail in anti-IGF2BP2. Overexpression of Snail partially reversed the regulatory effects of METTL3 on EMT-related gene expressions and metastatic abilities in NPC.
METTL3 is upregulated in NPC, which regulates EMT and metastasis in NPC cells through mA-modified Snail mRNA.
本研究旨在阐明METTL3通过对Snail进行甲基化修饰从而刺激上皮-间质转化（EMT），在加重鼻咽癌进展中的作用。
采用定量实时聚合酶链反应（qRT-PCR）检测48对鼻咽癌组织和癌旁组织中METTL3的差异表达。检测不同临床分期和转移状态的鼻咽癌患者中METTL3的水平。采用Kaplan-Meier法评估METTL3在鼻咽癌患者中的预后潜力。敲低METTL3后，检测SUNE-1细胞中Snail和EMT相关基因的表达变化以及侵袭和迁移能力。通过RNA结合蛋白免疫沉淀（RIP）试验验证Snail与METTL3和IGF2BP2之间的相互作用。最后，阐明METTL3/Snail调控环在影响鼻咽癌EMT和转移中的作用。
鼻咽癌组织中METTL3的表达高于癌旁组织。晚期或有淋巴结转移的鼻咽癌患者METTL3表达水平较高。Kaplan-Meier曲线显示，METTL3高表达的鼻咽癌患者预后较差。敲低METTL3可下调SUNE-1细胞中Snail和N-钙黏蛋白的蛋白水平，而上调E-钙黏蛋白的水平。同时，敲低METTL3可抑制鼻咽癌细胞的侵袭和迁移能力。RIP试验证实了Snail与METTL3之间的相互作用。此外，敲低METTL3降低了抗IGF2BP2中Snail的富集丰度。Snail的过表达部分逆转了METTL3对鼻咽癌EMT相关基因表达和转移能力的调控作用。
鼻咽癌中METTL3表达上调，其通过对Snail mRNA进行甲基化修饰来调节鼻咽癌细胞的EMT和转移。
