Wang Qiang, Chen Mengyuan, Tang Xiaofang
Department of Clinical Laboratory, The Second Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310005, People's Republic of China.
The Second Clinical Medical College, Zhejiang Chinese Medical University, Hangzhou, People's Republic of China.
Integr Cancer Ther. 2024 Jan-Dec;23:15347354241247223. doi: 10.1177/15347354241247223.
Luteolin, a common dietary flavonoid found in plants, has been shown to have anti-cancer properties. However, its exact mechanisms of action in non-small cell lung cancer (NSCLC) are still not fully understood, particularly its role in regulating broader genomic networks and specific gene targets. In this study, we aimed to elucidate the role of microRNAs (miRNAs) in NSCLC treated with luteolin, using A549 cells as a model system.
miRNA profiling was conducted on luteolin-treated A549 cells using Exiqon microarrays, with validation of selected miRNAs by qRT-PCR. Bioinformatic analysis identified the regulatory roles of miRNAs in biological processes and pathways following luteolin treatment. Computational algorithms were employed to identify potential target genes. A549 cells were transfected with miR-106a-5p mimic and inhibitor or their corresponding controls. The expression levels of 2 genes, twist basic helix-loop-helix transcription factor 1 (TWIST1) and matrix metallopeptidase 2 (MMP2), and cell migration were assessed.
miRNA profiling identified 341 miRNAs, with 18 exhibiting significantly altered expression ( < 0.05). Subsequent qRT-PCR analysis confirmed altered expression of 6 selected miRNAs. KEGG and GO analyses revealed significant alterations in pathways and biological processes crucial for tumor biology. TWIST1 and MMP2, which both contain conserved miR-106a-5p binding sites, exhibited an inverse correlation with the expression levels of miR-106a-5p. Dual-luciferase reporter assays confirmed TWIST1 and MMP2 as direct targets of miR-106a-5p. Luteolin treatment led to a reduction in A549 cell migration, and this reduction was further amplified by the overexpression of miR-106a-5p.
Luteolin inhibits A549 cell migration by modulating the miRNA landscape, shedding light on its mechanisms and laying the foundation for miRNA-based therapeutic approaches for NSCLC.
木犀草素是一种在植物中常见的膳食类黄酮，已被证明具有抗癌特性。然而，其在非小细胞肺癌（NSCLC）中的确切作用机制仍未完全阐明，尤其是其在调节更广泛的基因组网络和特定基因靶点方面的作用。在本研究中，我们旨在以A549细胞为模型系统，阐明木犀草素处理的NSCLC中微小RNA（miRNA）的作用。
使用Exiqon微阵列对木犀草素处理的A549细胞进行miRNA谱分析，并通过qRT-PCR对选定的miRNA进行验证。生物信息学分析确定了木犀草素处理后miRNA在生物学过程和通路中的调节作用。采用计算算法鉴定潜在的靶基因。用miR-106a-5p模拟物和抑制剂或其相应对照转染A549细胞。评估了2个基因，即扭曲基本螺旋-环-螺旋转录因子1（TWIST1）和基质金属肽酶2（MMP2）的表达水平以及细胞迁移情况。
miRNA谱分析鉴定出341种miRNA，其中18种表现出显著改变的表达（<0.05）。随后的qRT-PCR分析证实了6种选定miRNA的表达改变。KEGG和GO分析揭示了对肿瘤生物学至关重要的通路和生物学过程中的显著改变。TWIST1和MMP2均含有保守的miR-106a-5p结合位点，它们与miR-106a-5p的表达水平呈负相关。双荧光素酶报告基因测定证实TWIST1和MMP2是miR-106a-5p的直接靶标。木犀草素处理导致A549细胞迁移减少，而miR-106a-5p的过表达进一步增强了这种减少。
木犀草素通过调节miRNA格局抑制A549细胞迁移，揭示了其作用机制，并为基于miRNA的NSCLC治疗方法奠定了基础。
