RNA 甲基转移酶样 3 (METTL3) 负责 N 6-甲基腺苷 (m6A) 修饰过程中的甲基转移。这种表观遗传特征有助于 RNA 的结构和功能调节，因此可能促进肿瘤发生、肿瘤进展和细胞对抗癌治疗（化疗、放疗和免疫治疗）的反应。在头颈鳞状细胞癌（HNSCC）中，常用的化疗药物是顺铂。不幸的是，顺铂耐药仍然是肿瘤复发和患者死亡的主要原因。因此，本研究旨在探讨 METTL3 在 HNSCC 体外模型中对顺铂细胞反应的作用。对用 CRISPR-Cas9 系统建立的稳定 METTL3 敲低（sgMETTL3）的 HNSCC 细胞系（H103、FaDu 和 Detroit-562）进行处理顺铂的耐受血浆水平 (TPL) 为 0.5，TPL 为 1。此外，与对照（用对照 sgRNA 转导的细胞）相比，分析了细胞周期分布、细胞凋亡、CD44/CD133 表面标志物表达和细胞集落形成的能力。细胞周期分布和细胞凋亡的分析表明细胞百分比显着更高METTL3 敲低 1) 在 G2/S 阶段停滞，2) 与对照相比，表现为晚期凋亡或死亡。集落形成测定显示，在 FaDu 和 Detroit-562 METTL3 缺陷细胞中，单个细胞生长成集落的能力受到强化抑制，而在顺铂处理后，在 H103 METTL3 敲低细胞中观察到更高的集落数量。此外，METTL3 缺陷显着增加了所有研究细胞系中癌症干细胞标志物的表面表达。我们的研究结果强调了 METTL3 对细胞对顺铂反应的显着影响，表明其作为解决某些癌症病例中顺铂耐药性的有前景的治疗靶点的潜力HNSCC.版权所有 © 2024 Ostrowska、Rawłuszko-Wieczorek、Ostapowicz、Suchorska 和 Golusiński。

RNA methyltransferase-like 3 (METTL3) is responsible for methyl group transfer in the progression of N 6-methyladenosine (m6A) modification. This epigenetic feature contributes to the structural and functional regulation of RNA and consequently may promote tumorigenesis, tumor progression, and cellular response to anticancer treatment (chemo-, radio-, and immunotherapy). In head and neck squamous cell carcinoma (HNSCC), the commonly used chemotherapy is cisplatin. Unfortunately, cisplatin resistance is still a major cause of tumor relapse and patients' death. Thus, this study aimed to investigate the role of METTL3 on cellular response to cisplatin in HNSCC in vitro models.HNSCC cell lines (H103, FaDu, and Detroit-562) with stable METTL3 knockdown (sgMETTL3) established with CRISPR-Cas9 system were treated with 0.5 tolerable plasma level (TPL) and 1 TPL of cisplatin. Further, cell cycle distribution, apoptosis, CD44/CD133 surface marker expression, and cell's ability to colony formation were analyzed in comparison to controls (cells transduced with control sgRNA).The analyses of cell cycle distribution and apoptosis indicated a significantly higher percentage of cells with METTL3 knockdown 1) arrested in the G2/S phase and 2) characterized as a late apoptotic or death in comparison to control. The colony formation assay showed intensified inhibition of a single cell's ability to grow into a colony in FaDu and Detroit-562 METTL3-deficient cells, while a higher colony number was observed in H103 METTL3 knockdown cells after cisplatin treatment. Also, METTL3 deficiency significantly increased cancer stem cell markers' surface expression in all studied cell lines.Our findings highlight the significant influence of METTL3 on the cellular response to cisplatin, suggesting its potential as a promising therapeutic target for addressing cisplatin resistance in certain cases of HNSCC.Copyright © 2024 Ostrowska, Rawłuszko-Wieczorek, Ostapowicz, Suchorska and Golusiński.