Cardiothoracic Surgery, Puyang Oilfield General Hospital, China.
Cytokine. 2022 Aug;156:155922. doi: 10.1016/j.cyto.2022.155922. Epub 2022 Jun 1.
E2F8 can modulate development and progression of various cancers including cervical cancer, breast cancer and hepatocellular carcinoma. But its mechanism in lung adenocarcinoma (LUAD) remains underexplored. In this study, we conducted a series of experiments including qRT-PCR, western blot, CCK-8, scratch healing assay, Transwell, and flow cytometry. Through these assays, we confirmed the notable overexpression of E2F8 in LUAD and its promoting effects on LUAD cell proliferation, migration and invasion. Subsequently, microRNA-1-3p that was negatively associated with E2F8 expression was identified through bioinformatics analysis. qRT-PCR was then carried out for quantification of microRNA-1-3p expression, which displayed low microRNA-1-3p expression in LUAD cells. In addition, dual-luciferase reporter gene assay was utilized for validating the targeted relationship between microRNA-1-3p and E2F8. The results denoted that microRNA-1-3p could bind to the promoter region of E2F8. Finally, the results of rescue experiment revealed that microRNA-1-3p negatively modulated E2F8 level. It regulated NF-κB pathway to repress LUAD cell proliferative, migratory, and invasive properties, lead to cell cycle arrest in G0/G1 phase, and enhance cell apoptosis level. This study unraveled that microRNA-1-3p/E2F8 constrained LUAD malignant progression through NF-κB pathway, which may provide possible targets for LUAD diagnosis and treatment.
E2F8 可以调节包括宫颈癌、乳腺癌和肝癌在内的多种癌症的发展和进展。但其在肺腺癌 (LUAD) 中的机制仍未得到充分探索。在这项研究中，我们进行了一系列实验，包括 qRT-PCR、western blot、CCK-8、划痕愈合实验、Transwell 和流式细胞术。通过这些实验，我们证实了 E2F8 在 LUAD 中的显著过表达及其对 LUAD 细胞增殖、迁移和侵袭的促进作用。随后，通过生物信息学分析确定了与 E2F8 表达呈负相关的 microRNA-1-3p。然后通过 qRT-PCR 对 microRNA-1-3p 的表达进行定量，结果显示 LUAD 细胞中的 microRNA-1-3p 表达水平较低。此外，还利用双荧光素酶报告基因实验验证了 microRNA-1-3p 和 E2F8 之间的靶向关系。结果表明，microRNA-1-3p 可以结合到 E2F8 的启动子区域。最后，挽救实验的结果表明，microRNA-1-3p 负调控 E2F8 水平。它通过 NF-κB 通路调节 LUAD 细胞的增殖、迁移和侵袭特性，导致细胞周期停滞在 G0/G1 期，并增强细胞凋亡水平。这项研究揭示了 microRNA-1-3p/E2F8 通过 NF-κB 通路抑制 LUAD 恶性进展，这可能为 LUAD 的诊断和治疗提供可能的靶点。
