Su Yongxu, Hu Yanjia, Qu Binbin, Lei Rongchang, Guo Ge
Department of Oral and Maxilofacial Sugery, Changsha Stomatological Hospital, Changsha, 410004, Hunan, China.
Department of Oral and Maxilofacial Sugery, Xiangya Stomatological Hospital Central South University, Changsha, 410000, Hunan, China.
Mol Biotechnol. 2025 May;67(5):1867-1879. doi: 10.1007/s12033-024-01165-y. Epub 2024 May 14.
Oral squamous cell carcinoma (OSCC) is a common and highly lethal epithelial cancer. This study aimed to confirm the role of METTL3 in promoting OSCC and investigate its specific underlying mechanisms. Expression of the METTL3, YTH domain-containing family 2 (YTHDF2), and WEE1 were examined in normal oral epithelial cells and OSCC cells. Cell functions were examined after overexpressing WEE1 in OSCC cells. MeRIP-qPCR analysis was used to detect WEE1 m6A levels in HOK, SCC25, and CAL27 cells. WEE1 and its m6A levels were evaluated in OSCC cells by knocking down METTL3/YTHDF2, assessing the interaction between METTL3/YTHDF2 and WEE1. The impact of METTL3 and YTHDF2 downregulation on WEE1 mRNA stability was also investigated. The tumor weight and volume in a nude mouse model of OSCC after overexpression of WEE1 and YTHDF2 were measured. Expression of Ki-67 and WEE1 in OSCC tissue was detected using immunohistochemistry. Compared to normal oral epithelial cells, METTL3 and YTHDF2 were upregulated in OSCC cells, while WEE1 was downregulated, and there was a negative correlation between WEE1 and METTL3/YTHDF2 expression. WEE1 overexpression inhibited proliferation, invasion, and migration while promoting apoptosis in OSCC cells. METTL3 and YTHDF2 bound to WEE1 mRNA. METTL3/YTHDF2 knockdown increased WEE1 levels and WEE1 mRNA stability. METTL3 inhibition reduced WEE1 m6A levels. Inhibition of METTL3 weakened the interaction between YTHDF2 and WEE1 mRNA. In vivo, overexpression of WEE1 suppressed OSCC development, which was reversed by overexpression of YTHDF2. METTL3 facilitates the progression of OSCC through m6A-YTHDF2-dependent downregulation of WEE1.
口腔鳞状细胞癌（OSCC）是一种常见且致死率高的上皮性癌症。本研究旨在证实METTL3在促进OSCC中的作用，并探究其具体潜在机制。检测了正常口腔上皮细胞和OSCC细胞中METTL3、含YTH结构域家族2（YTHDF2）和WEE1的表达。在OSCC细胞中过表达WEE1后检测细胞功能。采用MeRIP-qPCR分析检测人正常口腔角质形成细胞（HOK）、SCC25和CAL27细胞中WEE1的m6A水平。通过敲低METTL3/YTHDF2评估OSCC细胞中WEE1及其m6A水平，检测METTL3/YTHDF2与WEE1之间的相互作用。还研究了METTL3和YTHDF2下调对WEE1 mRNA稳定性的影响。测量了过表达WEE1和YTHDF2后OSCC裸鼠模型中的肿瘤重量和体积。采用免疫组织化学检测OSCC组织中Ki-67和WEE1的表达。与正常口腔上皮细胞相比，OSCC细胞中METTL3和YTHDF2上调，而WEE1下调，且WEE1与METTL3/YTHDF2表达呈负相关。WEE1过表达抑制OSCC细胞的增殖、侵袭和迁移，同时促进细胞凋亡。METTL3和YTHDF2与WEE1 mRNA结合。敲低METTL3/YTHDF2可增加WEE1水平和WEE1 mRNA稳定性。抑制METTL3可降低WEE1的m6A水平。抑制METTL3可减弱YTHDF2与WEE1 mRNA之间的相互作用。在体内，WEE1过表达抑制OSCC发展，而YTHDF2过表达可逆转这一作用。METTL3通过m6A-YTHDF2依赖的WEE1下调促进OSCC进展。
