ZC3H13 通过 m6A 依赖性修饰减少喉鳞状细胞癌细胞中 DUOX1 介导的铁死亡。

ZC3H13 reduced DUOX1-mediated ferroptosis in laryngeal squamous cell carcinoma cells through m6A-dependent modification.

机构信息

Department of Otorhinolaryngology, The First People's Hospital of Fuyang Hangzhou, 311400 Hangzhou, Zhejiang Province, China.

出版信息

Tissue Cell. 2023 Oct;84:102187. doi: 10.1016/j.tice.2023.102187. Epub 2023 Jul 29.

DOI:10.1016/j.tice.2023.102187

PMID:37536262

Abstract

Laryngeal squamous cell carcinoma (LSCC) is the second most common head and neck cancer. To identify the link between ferroptosis and LSCC, we targeted the dual oxidase 1 (DUOX1) gene. This study aimed to reveal the intrinsic mechanism by which the DUOX1-zinc-finger CCCH domain-containing protein 13 (ZC3H13) ferroptosis axis affected the LSCC process. GEPIA was used to investigate the expression of DUOX1 in LSCC, and the expression levels of DUOX1 and ZC3H13 were manipulated by overexpression and RNA interference. MTT assay was used to detect cell proliferation. Chromatin immunoprecipitation (CHIP) detected the binding of DUOX1 and ZC3H13, and ROS assessment and intracellular Fe content determination were performed to examine the ferroptosis. MeRIP was used to analyze the m6A methylation of DUOX1. Ferroptosis-related proteins were detected by qRT-PCR. DUOX1 was found to be poorly expressed in LSCC cells, low DUOX1 level promoted LSCC cell proliferation, and low ZC3H13 level decreased LSCC cell proliferation. Besides, there was an interaction between DUOX1 and ZC3H13. DUOX1 could inhibit the expression levels of ferroptosis-related genes GPX4 and F1H1 in LSCC cells DUOX1 inhibited the expression levels of ROS and ferroptosis-related genes GPX4 and F1H1 and increased intracellular iron content in LSCC cells, but ZC3H13 reversed this phenomenon by inhibiting DUOX1 gene through m6A methylation modification. ZC3H13 reduced DUOX1-mediated ferroptosis in LSCC cells through m6A-dependent modification. The regulatory pathway of DUOX1 and ferroptosis are potential targets for designing diagnostic and combination therapeutic strategies for LSCC patients.

摘要

喉鳞状细胞癌（LSCC）是第二大常见的头颈部癌症。为了确定铁死亡与 LSCC 之间的联系，我们将目标锁定在双氧化酶 1（DUOX1）基因上。本研究旨在揭示 DUOX1-锌指 CCCH 结构域蛋白 13（ZC3H13）铁死亡轴影响 LSCC 过程的内在机制。GEPIA 用于研究 DUOX1 在 LSCC 中的表达，通过过表达和 RNA 干扰来操纵 DUOX1 和 ZC3H13 的表达水平。MTT 测定法用于检测细胞增殖。染色质免疫沉淀（CHIP）检测 DUOX1 和 ZC3H13 的结合，通过 ROS 评估和细胞内铁含量测定来检查铁死亡。MeRIP 用于分析 DUOX1 的 m6A 甲基化。通过 qRT-PCR 检测铁死亡相关蛋白。发现 DUOX1 在 LSCC 细胞中表达水平较低，低 DUOX1 水平促进 LSCC 细胞增殖，低 ZC3H13 水平降低 LSCC 细胞增殖。此外，DUOX1 和 ZC3H13 之间存在相互作用。DUOX1 可以抑制 LSCC 细胞中与铁死亡相关的基因 GPX4 和 F1H1 的表达水平。DUOX1 抑制 LSCC 细胞中 ROS 和铁死亡相关基因 GPX4 和 F1H1 的表达水平并增加细胞内铁含量，但 ZC3H13 通过 m6A 甲基化修饰抑制 DUOX1 基因，从而逆转这种现象。ZC3H13 通过 m6A 依赖性修饰降低 DUOX1 介导的 LSCC 细胞铁死亡。DUOX1 和铁死亡的调控途径是为 LSCC 患者设计诊断和联合治疗策略的潜在靶点。