Wang Man, Qin Yujiao, Ai Xiaoqi, Liu Xiuhua
Department of Respiratory Medicine, The First Affiliated Hospital of Jilin University, 1 Xinmin Street, Changchun, 130021, Jilin, China.
Mol Med. 2024 Dec 23;30(1):267. doi: 10.1186/s10020-024-01018-z.
Non-small cell lung cancer (NSCLC) is the predominant form of lung cancer, contributing significantly to global health and economic challenges. This study elucidated the role of RBM15 in NSCLC progression through its involvement in m6A modifications.
RBM15 levels in NSCLC tissues and cells were assessed via RT-qPCR and Western blotting. The impact of RBM15 knockdown on NSCLC proliferation, invasion, and migration was evaluated using CCK-8, colony formation, and Transwell assays. Expression levels of KLF1, TRIM13, and ANXA8 were determined by RT-qPCR and Western blot. m6A methylation levels were analyzed, while RIP and MeRIP assays were employed to explore the interaction between YTHDF1/YTHDF2/m6A and KLF1/TRIM13, as well as KLF1 binding to the ANXA8 promoter. The ubiquitination of ANXA8 was examined through ubiquitination assays. Xenograft and metastasis models were utilized to assess RBM15's role in vivo.
RBM15 was found to be overexpressed in NSCLC. Silencing RBM15 led to decreased cell proliferation, invasion, and migration of NSCLC cells. RBM15 upregulated KLF1 and downregulated TRIM13 via YTHDF1/YTHDF2, resulting in the promotion of ANXA8 expression. KLF1 overexpression or TRIM13 downregulation partially reversed the suppressive effects of RBM15 knockdown on NSCLC cell proliferation. ANXA8, upregulated in NSCLC, mitigated the inhibitory effects of RBM15 silencing on malignant behaviors. In vivo, RBM15 downregulation hindered NSCLC cell proliferation and metastasis by modulating the KLF1-TRIM13/ANXA8 axis.
RBM15-mediated m6A methylation enhances KLF1 expression and suppresses TRIM13 via YTHDF1/YTHDF2, thereby promoting ANXA8 and facilitating NSCLC progression. These findings provide novel insights and potential therapeutic targets for NSCLC treatment.
非小细胞肺癌（NSCLC）是肺癌的主要形式，对全球健康和经济构成重大挑战。本研究通过RBM15参与m6A修饰阐明了其在NSCLC进展中的作用。
通过RT-qPCR和蛋白质免疫印迹法评估NSCLC组织和细胞中RBM15的水平。使用CCK-8、集落形成和Transwell实验评估RBM15敲低对NSCLC增殖、侵袭和迁移的影响。通过RT-qPCR和蛋白质免疫印迹法测定KLF1、TRIM13和ANXA8的表达水平。分析m6A甲基化水平，同时采用RNA免疫沉淀（RIP）和甲基化RNA免疫沉淀（MeRIP）实验探究YTHDF1/YTHDF2/m6A与KLF1/TRIM13之间的相互作用，以及KLF1与ANXA8启动子的结合情况。通过泛素化实验检测ANXA8的泛素化。利用异种移植和转移模型评估RBM15在体内的作用。
发现RBM15在NSCLC中过表达。沉默RBM15导致NSCLC细胞的增殖、侵袭和迁移减少。RBM15通过YTHDF1/YTHDF2上调KLF1并下调TRIM13，从而促进ANXA8的表达。KLF1过表达或TRIM13下调部分逆转了RBM15敲低对NSCLC细胞增殖的抑制作用。在NSCLC中上调的ANXA8减轻了RBM15沉默对恶性行为的抑制作用。在体内，RBM15下调通过调节KLF1-TRIM13/ANXA8轴阻碍NSCLC细胞的增殖和转移。
RBM15介导的m6A甲基化通过YTHDF1/YTHDF2增强KLF1表达并抑制TRIM13，从而促进ANXA8表达并促进NSCLC进展。这些发现为NSCLC治疗提供了新的见解和潜在的治疗靶点。
