ZC3H13 的过表达通过 m6A 修饰介导的 IQGAP1 降解抑制甲状腺乳头状癌的生长。

Overexpressed ZC3H13 suppresses papillary thyroid carcinoma growth through m6A modification-mediated IQGAP1 degradation.

机构信息

Department of Thyroid Surgery, Hongjiaozhou Branch of the Second Affiliated Hospital of Nanchang University, Xuefu Avenue, Honggutan District, Nanchang 330006, Jiangxi, China.

出版信息

J Formos Med Assoc. 2023 Aug;122(8):738-746. doi: 10.1016/j.jfma.2022.12.019. Epub 2023 Feb 2.

DOI:10.1016/j.jfma.2022.12.019

PMID:36739231

Abstract

PURPOSE

The purpose of this study was to clarify the effect of ZC3H13 on the growth of papillary thyroid carcinoma (PTC).

METHODS

Firstly, we used qRT-PCR and Western blot to compare the difference in the expression of ZC3H13 between normal thyroid epithelial cells and PTC cell lines. Then, ZC3H13 overexpression/knockout thyroid cancer cells were constructed by lentivirus transfection, and the effects of overexpression of ZC3H13 on the proliferation, migration and invasion of PTC cells were detected by CCK8 and transwell experiments. Lastly, MeRIP-qPCR, RIP and o Actinomycin D were used to verify that ZC3H13 regulated the expression of downstream target gene IQGAP1 through m6A modification.

RESULTS

ZC3H13 expression was decreased in PTC cell lines BCPAP, KTC-1, k1, HTH83, and TPC-1. Proliferation, invasion, and migration of PTC cells were inhibited by overexpressed ZC3H13 but increased by knockdown of ZC3H13. IQGAP1 expression was suppressed by ZC3H13 overexpression but enhanced by ZC3H13 knockdown. In ZC3H13-overexpressed PTC cells, the m6A level of IQGAP1 mRNA was increased, and the IQGAP1 mRNA expression was decreased with the increasing time of Actinomycin D treatment. YTHDF2 enriched more IQGAP1 mRNA than IgG and knockdown of YTHDF2 reversed the effect of ZC3H13 overexpression on IQGAP1 mRNA stability. The xenograft tumor experiment in nude mice confirmed that the overexpression of ZC3H13 inhibited tumor growth, while overexpression of IQGAP1 could reverse the inhibitory effect of ZC3H13 overexpression on tumor growth.

CONCLUSION

ZC3H13 mediates IQGAP1 mRNA degradation by promoting m6A modification of IQGAP1 mRNA, this provides a prospective therapeutic target for PTC.

摘要

目的

本研究旨在阐明 ZC3H13 对甲状腺乳头状癌（PTC）生长的影响。

方法

首先，我们使用 qRT-PCR 和 Western blot 比较正常甲状腺上皮细胞和 PTC 细胞系中 ZC3H13 的表达差异。然后，通过慢病毒转染构建 ZC3H13 过表达/敲除甲状腺癌细胞，通过 CCK8 和 Transwell 实验检测 ZC3H13 过表达对 PTC 细胞增殖、迁移和侵袭的影响。最后，使用 MeRIP-qPCR、RIP 和 Actinomycin D 验证 ZC3H13 通过 m6A 修饰调控下游靶基因 IQGAP1 的表达。

结果

在 PTC 细胞系 BCPAP、KTC-1、k1、HTH83 和 TPC-1 中，ZC3H13 的表达降低。过表达 ZC3H13 可抑制 PTC 细胞的增殖、侵袭和迁移，而敲低 ZC3H13 则可促进其增殖、侵袭和迁移。ZC3H13 过表达可抑制 IQGAP1 的表达，而敲低 ZC3H13 则可增强 IQGAP1 的表达。在 ZC3H13 过表达的 PTC 细胞中，IQGAP1 mRNA 的 m6A 水平增加，随着 Actinomycin D 处理时间的增加，IQGAP1 mRNA 的表达减少。YTHDF2 比 IgG 更多地富集 IQGAP1 mRNA，而敲低 YTHDF2 则逆转了 ZC3H13 过表达对 IQGAP1 mRNA 稳定性的影响。裸鼠异种移植瘤实验证实，ZC3H13 的过表达抑制肿瘤生长，而过表达 IQGAP1 可逆转 ZC3H13 过表达对肿瘤生长的抑制作用。