Lu Zhiying, Chen Jing, Luo Chao
Department of Pediatrics, Wuhan Fourth Hospital, No.76, Jiefang Avenue, Qiaokou District, Wuhan, 430034, Hubei, China.
Cell Div. 2025 Apr 23;20(1):10. doi: 10.1186/s13008-025-00155-z.
Several studies have delineated that dysregulated N6-methyladenosine (m6A) regulators participate in glioma progression. The objective of this study is to investigate the mechanism of Wilms' tumor 1-associating protein (WTAP)-mediated m6A modification of long noncoding RNA (lncRNA) LINC00200 in glioma.
The LINC00200 expression in glioma was analyzed by qRT-PCR. The expressions of WTAP and Wnt/β-catenin pathway associated proteins were determined via qRT-PCR or western blotting. The levels of WTAP-mediated m6A modification of LINC00200 was ascertained by MeRIP-qPCR. Functionally, the effects of LINC00200 knockdown and the interaction of WTAP with LINC00200 on the glioma cell characteristics were examined by CCK8, colony formation, and transwell migration/invasion assays. In vivo experiments were performed to verify the effect of LINC00200 on tumor growth.
LINC00200 was overexpressed in glioma, and high LINC00200 level was related to higher-grade tumor. Moreover, its knockdown inhibited the malignant properties and expression of molecules related to Wnt/β-catenin pathway in glioma cell lines. In vivo, LINC00200 knockdown attenuated tumor growth. WTAP was also overexpressed in glioma tissues and demonstrated a positive association with LINC00200 expression. Furthermore, the relative enrichment of LINC00200 m6A was enhanced/reduced in a WTAP-dependent manner. Meanwhile, silencing LINC00200 partially reversed the malignant effects of WTAP overexpression in glioma.
These results demonstrate that WTAP-mediated m6A modification of LINC00200 promotes glioma progression by modulating Wnt/β-catenin pathway.
多项研究表明，失调的N6-甲基腺苷（m6A）调节因子参与神经胶质瘤进展。本研究旨在探讨威尔姆斯瘤1相关蛋白（WTAP）介导的长链非编码RNA（lncRNA）LINC00200的m6A修饰在神经胶质瘤中的作用机制。
采用qRT-PCR分析神经胶质瘤中LINC00200的表达。通过qRT-PCR或蛋白质免疫印迹法检测WTAP及Wnt/β-连环蛋白信号通路相关蛋白的表达。采用甲基化RNA免疫沉淀定量PCR（MeRIP-qPCR）确定WTAP介导的LINC00200的m6A修饰水平。功能上，通过CCK8、集落形成及Transwell迁移/侵袭实验检测LINC00200敲低以及WTAP与LINC00200相互作用对神经胶质瘤细胞特性的影响。进行体内实验以验证LINC00200对肿瘤生长的影响。
LINC00200在神经胶质瘤中过表达，且LINC00200高表达与高级别肿瘤相关。此外，敲低LINC00200可抑制神经胶质瘤细胞系的恶性特性及与Wnt/β-连环蛋白信号通路相关分子的表达。在体内，敲低LINC00200可减缓肿瘤生长。WTAP在神经胶质瘤组织中也过表达，且与LINC00200表达呈正相关。此外，LINC00200的m6A相对富集以WTAP依赖的方式增强/降低。同时，沉默LINC00200可部分逆转WTAP过表达对神经胶质瘤的恶性影响。
这些结果表明，WTAP介导的LINC00200的m6A修饰通过调节Wnt/β-连环蛋白信号通路促进神经胶质瘤进展。
