长链非编码RNA（lncRNA）的失调与非小细胞肺癌（NSCLC）的肿瘤发生和铁死亡有关。BBOX1反义RNA 1（BBOX1-AS1）在NSCLC中起着致癌驱动作用。在这里，我们旨在研究BBOX1-AS1在NSCLC进展和铁死亡中的调控效应和潜在机制。定量实时荧光定量聚合酶链反应（qRT-PCR）检测RNA表达，免疫印迹法测定蛋白质表达。细胞生长通过CCK-8和克隆形成实验评估。透过瞬移实验评估细胞入侵和迁移。RNA拉下和双荧光素酶报告基因检测法应用于验证miR-326与BBOX1-AS1或prominin 2（PROM2）之间的关系。通过异育瘤实验分析BBOX1-AS1在NSCLC肿瘤性的作用。沉默BBOX1-AS1或PROM2阻碍NSCLC细胞的生长、迁移和侵袭。沉默BBOX1-AS1诱导细胞凋亡和铁死亡。BBOX1-AS1上调PROM2的表达，而重新表达PROM2逆转了BBOX1-AS1减少对细胞恶性表型和铁死亡的影响。BBOX1-AS1通过海绵miR-326对PROM2的转录后修饰进行调控。miR-326被验证为BBOX1-AS1调控NSCLC细胞恶性表型和铁死亡的介导物质。此外，体内BBOX1-AS1缺陷导致异育瘤的抑制。综上，我们的研究定义了一个新的BBOX1-AS1/miR-326/PROM2轴对NSCLC恶性进展和铁死亡的调控作用，并为BBOX1-AS1在NSCLC治疗中的靶向应用提供了新证据。© 2023. 作者（们）已独家授权给Springer Science+Business Media, LLC，Springer Nature的一部分。

Dysregulation of long non-coding RNAs (lncRNAs) is associated with the tumorigenesis and ferroptosis of non-small cell lung cancer (NSCLC). BBOX1 antisense RNA 1 (BBOX1-AS1) functions as an oncogenic driver in NSCLC. Here, we aim to investigate the regulation effect and underlying mechanism of BBOX1-AS1 in NSCLC progression and ferroptosis. RNA expression was detected by quantitative real-time PCR (qRT-PCR), and protein expression was measured by immunoblotting. Cell growth was assessed by CCK-8 and colony formation assays. Transwell assay was applied to evaluate cell invasion and migration. RNA pull-down and dual-luciferase reporter assays were applied to verify the relationship between miR-326 and BBOX1-AS1 or prominin 2 (PROM2). The role of BBOX1-AS1 in NSCLC tumorigenicity was also analyzed by xenograft assays. Silencing BBOX1-AS1 or PROM2 impeded NSCLC cell growth, migration, and invasion. Silencing BBOX1-AS1 induced cell apoptosis and ferroptosis. BBOX1-AS1 up-regulated PROM2 expression, and re-expression of PROM2 reversed the effects of BBOX1-AS1 depletion on cell malignant phenotypes and ferroptosis. BBOX1-AS1 post-transcriptionally modulated PROM2 expression by sponging miR-326. MiR-326 was validated as a mediator of BBOX1-AS1 in regulating NSCLC cell malignant phenotypes and ferroptosis. Additionally, BBOX1-AS1 deficiency in vivo resulted in the suppression of xenograft tumor growth. Together, our study defines a novel BBOX1-AS1/miR-326/PROM2 axis in regulating NSCLC malignant progression and ferroptosis, offering new evidence for the oncogenic role of BBOX1-AS1 in NSCLC. These findings may provide a basis for the future usage of targeting BBOX1-AS1 in NSCLC treatment.© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.