Chen Xinming, Xu Xiaodong, Li Yifan, Shen Zhenya
Department of Cardiovascular Surgery, The First Affiliated Hospital and Institute for Cardiovascular Science, Soochow University, Suzhou, China.
Department of Thoracic Surgery, Affiliated Hospital of Nantong University, Nantong, China.
J Thorac Dis. 2025 Jun 30;17(6):3873-3885. doi: 10.21037/jtd-24-999. Epub 2025 Jun 3.
Dysregulated microRNAs (miRNAs) play a crucial role in the progression of non-small cell lung cancer (NSCLC) through post-transcriptional gene regulation. Although miR-26b has been implicated in tumor suppression, its epigenetic regulation and functional targets in NSCLC remain poorly understood. DNA methylation is a well-established modulator of miRNA expression; however, the specific role of miR-26b hypermethylation and its interaction with oncogenic pathways, particularly reticulocalbin-1 (RCN1), have not been fully elucidated. This study aims to clarify the miR-26b/RCN1 regulatory axis in NSCLC pathogenesis. By establishing this mechanism, we seek to identify novel therapeutic strategies and improve patient outcomes.
We first quantified the expression levels of miR-26b in NSCLC cells using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and then evaluated its impact on cell proliferation, migration, and invasion using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), colony formation, and transwell assays. DNA methylation status was assessed by methylation-specific PCR (MSP) and bisulfite sequencing PCR (BSP). Western blotting and luciferase reporter assays were conducted to confirm the direct binding of miR-26b to the RCN1 3'-untranslated region (3'UTR). Additionally, we examined the effects of the miR-26b/RCN1 axis on NSCLC cell behaviors .
Our findings demonstrated that RCN1 was a direct target of miR-26b, and miR-26b expression was downregulated due to DNA hypermethylation. Luciferase assays confirmed that miR-26b suppressed RCN1 expression by targeting its 3'UTR. Moreover, overexpression of miR-26b significantly inhibited NSCLC cell proliferation, invasion, and migration by targeting RCN1.
Our results indicate that miR-26b plays a critical role in NSCLC progression, suggesting its potential as a therapeutic biomarker for NSCLC patients.
失调的微小RNA（miRNA）通过转录后基因调控在非小细胞肺癌（NSCLC）的进展中起关键作用。尽管miR-26b与肿瘤抑制有关，但其在NSCLC中的表观遗传调控和功能靶点仍知之甚少。DNA甲基化是一种公认的miRNA表达调节剂；然而，miR-26b高甲基化的具体作用及其与致癌途径，特别是网织钙结合蛋白-1（RCN1）的相互作用尚未完全阐明。本研究旨在阐明NSCLC发病机制中的miR-26b/RCN1调控轴。通过建立这一机制，我们试图确定新的治疗策略并改善患者预后。
我们首先使用定量逆转录聚合酶链反应（qRT-PCR）定量NSCLC细胞中miR-26b的表达水平，然后使用3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）、集落形成和Transwell试验评估其对细胞增殖、迁移和侵袭的影响。通过甲基化特异性PCR（MSP）和亚硫酸氢盐测序PCR（BSP）评估DNA甲基化状态。进行蛋白质免疫印迹和荧光素酶报告基因试验以确认miR-26b与RCN1 3'非翻译区（3'UTR）的直接结合。此外，我们研究了miR-26b/RCN1轴对NSCLC细胞行为的影响。
我们的研究结果表明，RCN1是miR-26b的直接靶点，并且由于DNA高甲基化，miR-26b的表达下调。荧光素酶试验证实，miR-26b通过靶向其3'UTR抑制RCN1表达。此外，miR-26b的过表达通过靶向RCN1显著抑制NSCLC细胞的增殖、侵袭和迁移。
我们的结果表明，miR-26b在NSCLC进展中起关键作用，表明其作为NSCLC患者治疗生物标志物的潜力。
