YTHDF2 通过激活 DDIT4/AKT/mTOR 信号通路促进间变性甲状腺癌的进展。

YTHDF2 promotes anaplastic thyroid cancer progression by activating the DDIT4/AKT/mTOR signaling pathway.

机构信息

Department of Thyroid and Hernia Surgery, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Southern Medical University, Guangzhou, Guangdong, 510080, China.

Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China.

出版信息

Biol Direct. 2024 Nov 26;19(1):122. doi: 10.1186/s13062-024-00566-y.

DOI:10.1186/s13062-024-00566-y

PMID:39593172

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11600618/

Abstract

BACKGROUND

RNA methylation, an important reversible post-transcriptional modification in eukaryotes, has emerged as a prevalent epigenetic alteration. However, the role of the m6A reader YTH domain family 2 (YTHDF2) has not been reported in anaplastic thyroid cancer (ATC) and its biological mechanism is unclear.

METHODS

The relationship between YTHDF2 expression and ATC was determined using data sets and tissue samples. A range of analytical techniques were employed to investigate the regulatory mechanism of YTHDF2 in ATC, including bioinformatics analysis, m6A dot-blot analysis, methylated RNA immunoprecipitation sequencing (MeRIP-seq), RNA immunoprecipitation (RIP) assays, RNA sequencing, RNA stability assays and dual luciferase reporter gene assays. In vitro and in vivo assays were also conducted to determine the contribution of YTHDF2 to ATC development.

RESULTS

YTHDF2 expression was significantly increased in ATC. The comprehensive in vitro and in vivo experiments demonstrated that YTHDF2 knockdown significantly attenuated ATC proliferation, invasion, migration, and apoptosis promotion, whereas YTHDF2 overexpression yielded the opposite trend. Mechanistically, RNA-seq, MeRIP-seq and RIP-seq analysis, and molecular biology experiments demonstrated that YTHDF2 accelerated the degradation of DNA damage-inducible transcript 4 or regulated in DNA damage and development 1 (DDIT4, or REDD1) mRNA in an m6A-dependent manner, which in turn activated the AKT/mTOR signaling pathway and induced activation of epithelial-mesenchymal transition (EMT), thereby promoting ATC tumor progression.

CONCLUSIONS

This study is the first to demonstrate that elevated YTHDF2 expression levels suppress DDIT4 expression in an m6A-dependent manner and activate the AKT/mTOR signaling pathway, thereby promoting ATC progression. YTHDF2 plays a pivotal role in ATC progression, and it may serve as a promising therapeutic target in the future.

摘要

背景

RNA 甲基化是真核生物中一种重要的可逆转录后修饰，已成为一种普遍的表观遗传改变。然而，m6A 阅读器 YTH 结构域家族 2（YTHDF2）在间变性甲状腺癌（ATC）中的作用尚未报道，其生物学机制尚不清楚。

方法

使用数据集和组织样本确定 YTHDF2 表达与 ATC 的关系。采用一系列分析技术研究 YTHDF2 在 ATC 中的调控机制，包括生物信息学分析、m6A 点印迹分析、甲基化 RNA 免疫沉淀测序（MeRIP-seq）、RNA 免疫沉淀（RIP）测定、RNA 测序、RNA 稳定性测定和双荧光素酶报告基因测定。还进行了体外和体内实验，以确定 YTHDF2 对 ATC 发展的贡献。

结果

YTHDF2 在 ATC 中表达显著增加。全面的体外和体内实验表明，YTHDF2 敲低显著抑制 ATC 的增殖、侵袭、迁移和促进凋亡，而 YTHDF2 过表达则产生相反的趋势。机制上，RNA-seq、MeRIP-seq 和 RIP-seq 分析以及分子生物学实验表明，YTHDF2 以 m6A 依赖的方式加速 DNA 损伤诱导转录物 4 或 DNA 损伤和发育调节蛋白 1（DDIT4，或 REDD1）mRNA 的降解，进而激活 AKT/mTOR 信号通路，并诱导上皮-间充质转化（EMT）的激活，从而促进 ATC 肿瘤的进展。