Cardiothoracic Surgery Department, The First People's Hospital of Fuyang, No. 429, Beihuan Road, Fuchun Street, Fuyang District, Hangzhou, 311400, Zhejiang, China.
Mol Biotechnol. 2022 Aug;64(8):852-860. doi: 10.1007/s12033-022-00465-5. Epub 2022 Feb 18.
Lung adenocarcinoma (LUAD) is the most frequent histological subtype of non-small cell lung cancer. Cyclin B1 (CCNB1) is the vital initiator and controller of mitosis. Studies have indicated that CCNB1 overexpression is closely associated with cell proliferation and tumorigenesis in many cancers. Thus, discovery of molecular mechanism of CCNB1 in LUAD is conducive to developing new diagnostic or therapeutic targets for LUAD. We acquired mature miRNA and mRNA expression information of LUAD from TCGA database, as well as related clinical data. CCNB1 expression in normal and LUAD tissue was analyzed. Relationship between CCNB1 and patient's survival and clinical stage was analyzed. Upstream regulatory gene miRNA of CCNB1 was predicted. qRT-PCR and western blot examined expression levels of CCNB1 and miR-139-5p in cells. CCK-8 tested cell proliferation. Scratch healing and Transwell determined cell migration and invasion. Flow cytometry analyzed the cell cycle. Dual-luciferase verified targeting relationship between the two genes. Compared to controls, CCNB1 expression was prominently high in LUAD patient samples, and associated with advanced tumor stages and shorter overall survival. MiR-139-5p expressed an evidently negative correlation with CCNB1 and was predicted to target CCNB1. MiR-139-5p mimics reduced CCNB1 mRNA and protein expression, and suppressed luciferase activity in a target-specific manner, as confirmed by a control construct with a mutated miR-139-5p binding site. CCNB1 overexpression fostered progression of LUAD cells. Mechanistically, miR-139-5p might negatively regulate CCNB1 in LUAD, thereby suppressing cell proliferation, migration, invasion and cell cycle.
肺腺癌（LUAD）是非小细胞肺癌中最常见的组织学亚型。细胞周期蛋白 B1（CCNB1）是有丝分裂的重要启动子和控制器。研究表明，CCNB1 过表达与许多癌症中的细胞增殖和肿瘤发生密切相关。因此，发现 LUAD 中 CCNB1 的分子机制有利于为 LUAD 开发新的诊断或治疗靶点。我们从 TCGA 数据库中获得了 LUAD 的成熟 miRNA 和 mRNA 表达信息以及相关的临床数据。分析了正常和 LUAD 组织中 CCNB1 的表达。分析了 CCNB1 与患者生存和临床分期的关系。预测了 CCNB1 的上游调控基因 miRNA。qRT-PCR 和 Western blot 检测了细胞中 CCNB1 和 miR-139-5p 的表达水平。CCK-8 检测细胞增殖。划痕愈合和 Transwell 测定细胞迁移和侵袭。流式细胞术分析细胞周期。双荧光素酶报告基因验证了两个基因之间的靶向关系。与对照组相比，LUAD 患者样本中 CCNB1 的表达明显升高，且与晚期肿瘤分期和总生存期缩短相关。miR-139-5p 与 CCNB1 表达呈明显负相关，并被预测为靶向 CCNB1。miR-139-5p 模拟物降低了 CCNB1 的 mRNA 和蛋白表达，并以靶特异性方式抑制了荧光素酶活性，通过具有突变的 miR-139-5p 结合位点的对照构建体得到证实。CCNB1 的过表达促进了 LUAD 细胞的进展。在机制上，miR-139-5p 可能在 LUAD 中负调控 CCNB1，从而抑制细胞增殖、迁移、侵袭和细胞周期。
