Department of Oncology, The People's Hospital of Shiyan, Shiyan, Hubei, China.
Department of Oncology, The Shiyan Hospital of Traditional Chinese, Shiyan, Hubei, China.
J Cell Biochem. 2020 Feb;121(2):1675-1689. doi: 10.1002/jcb.29402. Epub 2019 Oct 8.
The bioactivity of microRNA-1827 (miR-1827) in lung adenocarcinoma cells would be explored. The expression level of gene and miR-1827 in 76 pairs of lung adenocarcinoma tissues and adjacent counterparts were analyzed by a quantitative real-time polymerase chain reaction. Primary lung adenocarcinoma cells were derived from patients' tissues. These cells were treated with miR-1827 agomir to mimic the upregulation of endogenous miR-1827. The malignant degree of lung adenocarcinoma cells in vitro was evaluated by cell proliferation, colony formation, transwell invasion, and apoptosis assays. Western blot analysis was used to observe the transition of lung adenocarcinoma cells from epithelial-to-mesenchymal. Target genes of miR-1827 were predicted by bioinformatics analysis. In addition, the interaction between miR-1827 and candidate messenger RNAs was verified by dual-luciferase reporter assay and AGO2-RNA immunoprecipitation. Besides, the effect of miR-1827 on tumors was verified by in vivo experiments. Transient gene overexpression was achieved by plasmids transfection. In this study, we found that the expression of miR-1827 was downregulated in lung adenocarcinoma, and its low expression was significantly correlated with the progression of lung adenocarcinoma and poor prognosis of patients. miR-1827 overexpression remarkably reduced the malignancy of primary lung adenocarcinoma cells in vitro. MYC and FAM83F were identified as two targeted genes of miR-1827 in lung adenocarcinoma cells. The levels of these two genes were upregulated in lung adenocarcinoma, and their high expression was significantly associated with the progression of lung adenocarcinoma and poor prognosis of patients. Overexpression of MYC or FAM83F attenuated the effects of miR-1827 on primary lung adenocarcinoma cells in vitro. In addition, in vivo experiments showed that miR-1827 inhibited tumor growth by reducing the levels of MYC and FAM83F. In conclusion, miR-1827 might repress the development of lung adenocarcinoma by targeting oncogenic genes MYC and FAM83F.
本文旨在探讨 microRNA-1827（miR-1827）在肺腺癌细胞中的生物活性。通过实时定量聚合酶链反应分析了 76 对肺腺癌组织及其相邻组织中基因和 miR-1827 的表达水平。原代肺腺癌细胞源自患者组织。这些细胞用 miR-1827 agomir 处理以模拟内源性 miR-1827 的上调。通过细胞增殖、集落形成、Transwell 侵袭和凋亡测定评估肺腺癌细胞在体外的恶性程度。Western blot 分析用于观察肺腺癌细胞从上皮到间充质的转变。通过生物信息学分析预测 miR-1827 的靶基因。此外，通过双荧光素酶报告基因测定和 AGO2-RNA 免疫沉淀验证了 miR-1827 与候选信使 RNA 之间的相互作用。此外，通过体内实验验证了 miR-1827 对肿瘤的影响。通过质粒转染实现瞬时基因过表达。在本研究中，我们发现 miR-1827 在肺腺癌中表达下调，其低表达与肺腺癌的进展和患者预后不良显著相关。miR-1827 过表达显著降低了原代肺腺癌细胞在体外的恶性程度。在肺腺癌细胞中鉴定出 MYC 和 FAM83F 为 miR-1827 的两个靶基因。这两个基因在肺腺癌中的表达上调，其高表达与肺腺癌的进展和患者预后不良显著相关。过表达 MYC 或 FAM83F 减弱了 miR-1827 对原代肺腺癌细胞体外的作用。此外，体内实验表明 miR-1827 通过降低 MYC 和 FAM83F 的水平抑制肿瘤生长。综上所述，miR-1827 可能通过靶向致癌基因 MYC 和 FAM83F 抑制肺腺癌的发展。
