Department of Pharmacy, Shanghai University of Medicine & Health Sciences Affiliated Sixth People's Hospital South Campus, Shanghai, China.
Fengxian Hospital and School of Pharmaceutical Sciences, Southern Medical University, Shanghai, China.
Cancer Gene Ther. 2024 Nov;31(11):1721-1733. doi: 10.1038/s41417-024-00830-3. Epub 2024 Sep 19.
Circular RNA (circRNA) has been demonstrated to play a pivotal role in tumor development. This study aimed to investigate the regulatory mechanism of circMBOAT2 in non-small cell lung cancer (NSCLC) and its association with tumor growth induced by chronic stress. We constructed stably transfected A549 and H1299 cell lines with circMBOAT2 overexpression and knockdown. Colony formation, scratch healing, Transwell and CCK-8 assays were conducted to evaluate the effects of circMBOAT2 in the presence or absence of norepinephrine (NE) treatment on the proliferation, migration, and invasion of NSCLC cells, respectively. Additionally, A chronic unpredictable mild stress (CUMS)-induced depression with heterotopic transplantation LLC and injection of antisense oligonucleotides (ASOs) targeting circMBOAT2 mouse model was established to evaluate the effect of chronic stress on tumorigenesis via circMBOAT2. Moreover, we investigated the regulatory effect of CCCTC binding factor (CTCF) on circMBOAT2 expression through in vivo and in vitro silencing of CTCF. Our results revealed a significant upregulation of circMBOAT2 in NSCLC cell lines and tumor tissues. circMBOAT2 knockdown inhibited the proliferation, migration, and invasion of NSCLC cells, while NE treatment reversed the cell suppression effect caused by circMBOAT2 knockdown. Notably, CUMS promoted tumor growth, while silencing circMBOAT2 inhibited tumor growth in vivo. Furthermore, we identified CTCF as the upstream regulator of circMBOAT2, which exhibited upregulation in NSCLC cells and tissues. Knockdown of CTCF reversed the promotional effect of CUMS on circMBOAT2 expression and tumor growth. Our findings provide evidence that CTCF mediates chronic stress in promoting of NSCLC progression through circMBOAT2. circMBOAT2 may serve as a potential biomarker and therapeutic target for NSCLC as well as the treatment of comorbid depression in NSCLC patients.
环状 RNA（circRNA）已被证明在肿瘤发展中发挥关键作用。本研究旨在探讨 circMBOAT2 在非小细胞肺癌（NSCLC）中的调控机制及其与慢性应激诱导的肿瘤生长的关系。我们构建了稳定转染的 A549 和 H1299 细胞系，过表达和敲低 circMBOAT2。进行集落形成、划痕愈合、Transwell 和 CCK-8 测定，以评估在有无去甲肾上腺素（NE）处理的情况下，circMBOAT2 对 NSCLC 细胞增殖、迁移和侵袭的影响。此外，建立了慢性不可预测轻度应激（CUMS）诱导的抑郁合并异位移植 LLC 和注射针对 circMBOAT2 的反义寡核苷酸（ASO）的小鼠模型，以评估慢性应激通过 circMBOAT2 对肿瘤发生的影响。此外，我们通过体内和体外沉默 CTCF 研究了 CTCF 对 circMBOAT2 表达的调节作用。我们的结果显示，circMBOAT2 在 NSCLC 细胞系和肿瘤组织中显著上调。circMBOAT2 敲低抑制 NSCLC 细胞的增殖、迁移和侵袭，而 NE 处理逆转了 circMBOAT2 敲低引起的细胞抑制作用。值得注意的是，CUMS 促进肿瘤生长，而沉默 circMBOAT2 则抑制体内肿瘤生长。此外，我们确定 CTCF 是 circMBOAT2 的上游调节因子，其在 NSCLC 细胞和组织中上调。沉默 CTCF 逆转了 CUMS 对 circMBOAT2 表达和肿瘤生长的促进作用。我们的研究结果表明，CTCF 通过 circMBOAT2 介导慢性应激促进 NSCLC 的进展。circMBOAT2 可能作为 NSCLC 的潜在生物标志物和治疗靶点，以及治疗 NSCLC 患者共病抑郁。
