Zhong Xuesong, Wang Sixue, Yang Xiaoli, Yang Xi, Zhou Linchang
Department of Urology, People's Hospital of Chuxiong Yi Autonomous Prefecture, Chuxiong City, China.
Department of Reproductive, People's Hospital of Chuxiong Yi Autonomous Prefecture, Chuxiong City, China.
Transl Androl Urol. 2024 Jul 31;13(7):1145-1163. doi: 10.21037/tau-23-682. Epub 2024 Jul 16.
Methyltransferase-like (METTL) plays an important role in various biological processes, but its role in prostate cancer (PCa) is still unclear. This study aimed to explore the mechanism by which methyltransferase-like 14 (METTL14) inhibits the physiological activity of PCa cells by increasing the N6-methyladenosine (m6A) modification of cyclin-dependent kinase 4 (CDK4).
Clinical samples were collected for bioinformatics analysis. A PCa mouse model was constructed. Cell counting kit-8 (CCK-8), flow cytometry, colony formation assays, scratch assays, Transwell assays, real-time quantitative polymerase chain reaction (RT-qPCR), immunofluorescence and western blotting were used to detect the corresponding indicators.
METTL14 was found to be beneficial to inhibit the proliferation, invasion, and migration of PCa cells. When the m6A RNA increased, the half-life of CDK4 mRNA decreased after oe-METTL14 (overexpression of METTL14). Overexpression of CDK4 reversed the effect of oe-METTL14. Coimmunoprecipitation experiments revealed there were interactions between CDK4 and forkhead box M1 (FOXM1). Transfection of si-CDK4 was similar to transfection of oe-METTL14. After transfection with oe-FOXM1, the invasion and migration ability of cells increased, and cell apoptosis decreased. After transfection with si-FOXM1 alone, autophagy related 7 (ATG7) expression was significantly downregulated, and autophagy levels were reduced. The overexpression of ATG7 reversed the effect of si-FOXM1. The tumor volume and weight of the oe-METTL14 group mice were significantly reduced, and tumor proliferation was decreased in comparison to untreated tumor-bearing mice.
METTL14 inhibits the invasion and migration of PCa cells and induces cell apoptosis by inhibiting CDK4 stability and FOXM1/ATG7-mediated autophagy.
类甲基转移酶（METTL）在多种生物学过程中发挥重要作用，但其在前列腺癌（PCa）中的作用仍不清楚。本研究旨在探讨类甲基转移酶14（METTL14）通过增加细胞周期蛋白依赖性激酶4（CDK4）的N6-甲基腺苷（m6A）修饰来抑制PCa细胞生理活性的机制。
收集临床样本进行生物信息学分析。构建PCa小鼠模型。使用细胞计数试剂盒-8（CCK-8）、流式细胞术、集落形成试验、划痕试验、Transwell试验、实时定量聚合酶链反应（RT-qPCR）、免疫荧光和蛋白质印迹法检测相应指标。
发现METTL14有利于抑制PCa细胞的增殖、侵袭和迁移。当m6A RNA增加时，过表达METTL14（oe-METTL14）后CDK4 mRNA的半衰期缩短。CDK4的过表达逆转了oe-METTL14的作用。免疫共沉淀实验表明CDK4与叉头框M1（FOXM1）之间存在相互作用。转染si-CDK4的效果与转染oe-METTL14相似。转染oe-FOXM1后，细胞的侵袭和迁移能力增强，细胞凋亡减少。单独转染si-FOXM1后，自噬相关蛋白7（ATG7）的表达显著下调，自噬水平降低。ATG7的过表达逆转了si-FOXM1的作用。与未处理的荷瘤小鼠相比，oe-METTL14组小鼠的肿瘤体积和重量显著减小，肿瘤增殖降低。
METTL14通过抑制CDK4稳定性以及FOXM1/ATG7介导的自噬来抑制PCa细胞的侵袭和迁移并诱导细胞凋亡。
