PDF(Pubmed)
Abstract：
BACKGROUND: The potential involvement of circular RNAs (circRNAs) and N6-methyladenosine (m6A) modification in the progression of Wilms tumor (WT) has not been fully elucidated. This study investigates the regulatory mechanisms and clinical significance of m6A-modified circMARK2 and its role in WT progression.
METHODS: We identified dysregulated circRNAs through deep sequencing and validated their expression by qRT-PCR in WT tissues. The biological functions of circMARK2 were assessed using clone formation, transwell migration, and orthotopic animal models. To dissect the underlying mechanisms, we employed RNA immunoprecipitation, RNA pull-down, dual-luciferase reporter assays, Western blotting, and immunofluorescence and immunohistochemical staining.
RESULTS: CircMARK2, upregulated in WT tissues, was found to be m6A-modified and promoted cytoplasmic export. It facilitated WT progression by stabilizing LIN28B mRNA through the circMARK2/IGF2BP2 interaction. In vitro and in vivo studies demonstrated that circMARK2 enhances the malignant behavior of WT cells. Clinically, higher circMARK2 levels in tumor tissues of WT patients were linked to increased tumor aggressiveness and reduced survival rates.
CONCLUSIONS: Our study provides the first comprehensive evidence that m6A-modified circMARK2 contributes to WT progression by enhancing LIN28B mRNA stability, promoting cellular aggressiveness. CircMARK2 emerges as a potential biomarker for prognosis and a promising target for therapeutic intervention in WT, underscoring the clinical relevance of m6A modification in pediatric renal cancer.
METHODS: We identified dysregulated circRNAs through deep sequencing and validated their expression by qRT-PCR in WT tissues. The biological functions of circMARK2 were assessed using clone formation, transwell migration, and orthotopic animal models. To dissect the underlying mechanisms, we employed RNA immunoprecipitation, RNA pull-down, dual-luciferase reporter assays, Western blotting, and immunofluorescence and immunohistochemical staining.
RESULTS: CircMARK2, upregulated in WT tissues, was found to be m6A-modified and promoted cytoplasmic export. It facilitated WT progression by stabilizing LIN28B mRNA through the circMARK2/IGF2BP2 interaction. In vitro and in vivo studies demonstrated that circMARK2 enhances the malignant behavior of WT cells. Clinically, higher circMARK2 levels in tumor tissues of WT patients were linked to increased tumor aggressiveness and reduced survival rates.
CONCLUSIONS: Our study provides the first comprehensive evidence that m6A-modified circMARK2 contributes to WT progression by enhancing LIN28B mRNA stability, promoting cellular aggressiveness. CircMARK2 emerges as a potential biomarker for prognosis and a promising target for therapeutic intervention in WT, underscoring the clinical relevance of m6A modification in pediatric renal cancer.
摘要：
背景：环状RNA（circRNAs）和N6-甲基腺苷（m6A）修饰在Wilms肿瘤（WT）进展中的潜在参与尚未完全阐明。本研究探讨了m6A修饰的circRMARK2的调节机制和临床意义及其在WT进展中的作用。
方法：我们通过深度测序鉴定了失调的circRNAs，并通过qRT-PCR验证了它们在WT组织中的表达。使用克隆形成评估circMARK2的生物学功能，Transwell迁移,和原位动物模型。为了剖析潜在的机制，我们用RNA免疫沉淀,RNA下拉，双荧光素酶报告分析，西方印迹,免疫荧光和免疫组织化学染色。
结果：在WT组织中上调的CircMARK2，被发现是m6A修饰的并促进了细胞质输出。它通过circMARK2/IGF2BP2相互作用稳定LIN28BmRNA来促进WT进展。体外和体内研究表明circMARK2增强WT细胞的恶性行为。临床上,WT患者肿瘤组织中较高的circMARK2水平与肿瘤侵袭性增加和生存率降低相关.
结论：我们的研究提供了第一个全面的证据，表明m6A修饰的circleMARK2通过增强LIN28BmRNA稳定性促进WT进展，促进细胞侵略性。CircMARK2成为WT预后的潜在生物标志物和有希望的治疗干预目标，强调m6A修饰在小儿肾癌中的临床意义。
方法：我们通过深度测序鉴定了失调的circRNAs，并通过qRT-PCR验证了它们在WT组织中的表达。使用克隆形成评估circMARK2的生物学功能，Transwell迁移,和原位动物模型。为了剖析潜在的机制，我们用RNA免疫沉淀,RNA下拉，双荧光素酶报告分析，西方印迹,免疫荧光和免疫组织化学染色。
结果：在WT组织中上调的CircMARK2，被发现是m6A修饰的并促进了细胞质输出。它通过circMARK2/IGF2BP2相互作用稳定LIN28BmRNA来促进WT进展。体外和体内研究表明circMARK2增强WT细胞的恶性行为。临床上,WT患者肿瘤组织中较高的circMARK2水平与肿瘤侵袭性增加和生存率降低相关.
结论：我们的研究提供了第一个全面的证据，表明m6A修饰的circleMARK2通过增强LIN28BmRNA稳定性促进WT进展，促进细胞侵略性。CircMARK2成为WT预后的潜在生物标志物和有希望的治疗干预目标，强调m6A修饰在小儿肾癌中的临床意义。
