■甲基转移酶(METTL)在各种生物过程中起着重要作用,但其在前列腺癌(PCa)中的作用仍不清楚。本研究旨在探讨甲基转移酶样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)修饰抑制PCa细胞生理活性的机制。
■收集临床样品用于生物信息学分析。构建PCa小鼠模型。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分析,实时定量聚合酶链反应(RT-qPCR),免疫荧光和免疫印迹检测相应指标。
■METTL14被发现有利于抑制增殖,入侵,和PCa细胞的迁移。当m6ARNA增加时,Oe-METTL14(过表达METTL14)后,CDK4mRNA的半衰期降低。CDK4的过表达逆转了oe-METTL14的作用。免疫共沉淀实验显示CDK4和叉头盒M1(FOXM1)之间存在相互作用。si-CDK4的转染与oe-METTL14的转染相似。转染oe-FOXM1后,细胞的侵袭和迁移能力增强,细胞凋亡减少。用si-FOXM1单独转染后,自噬相关蛋白7(ATG7)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。Oe-METTL14组小鼠的肿瘤体积和体重显著降低,与未经治疗的荷瘤小鼠相比,肿瘤增殖降低。
■METTL14通过抑制CDK4稳定性和FOXM1/ATG7介导的自噬,抑制PCa细胞的侵袭和迁移并诱导细胞凋亡。
UNASSIGNED: 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).
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: METTL14 inhibits the invasion and migration of PCa cells and induces cell apoptosis by inhibiting CDK4 stability and FOXM1/ATG7-mediated autophagy.