Department of Pathology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region 530021, P.R. China.
Department of Medical Oncology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region 530021, P.R. China.
Mol Med Rep. 2017 Dec;16(6):8781-8792. doi: 10.3892/mmr.2017.7713. Epub 2017 Oct 4.
Our previous study demonstrated that the expression of miR‑146a‑5p was downregulated in non‑small cell lung cancer (NSCLC) tissue, which affected the progression and prognosis of patients with NSCLC. Thus, the present study was conducted to investigate the functional mechanism of miR‑146a‑5p in tumorigenesis and angiogenesis in NSCLC. Following the construction of a H460 NSCLC cell line in which miR‑146a‑5p was overexpressed via lentivirus transduction, the NSCLC chick embryo chorioallantoic membrane (CAM) model was established by transplanting miR‑146a‑5p‑overexpressing NSCLC cells into the CAM. Then, the size of the neoplasms within the CAM was measured, the vessel ratio was calculated, and the cellular morphology, metastasis and inflammation of tumor cell was observed using hematoxylin and eosin staining. The target genes of miR‑146a‑5p were predicted by 12 online software programs; these genes were then subjected to Gene Ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway annotations using the Database for Annotation, Visualization and Integrated Discovery 6.7 as well as constructed into a protein interaction network using protein‑protein interaction from Search Tool for the Retrieval of Interacting Genes/Proteins. The xenograft tumor size and angiogenesis conditions of the miR‑146a‑5p‑overexpressing group (volume 6.340±0.066 mm3, vessel ratio 9.326±0.083) was obviously restricted (P<0.001) when compared with the low expression group (volume 30.13±0.06 mm3, vessel ratio 16.94±0.11). In addition, marked necrosis along with inflammatory cell infiltration was observed with the HE‑stained slices from the miR‑146a‑5p low expression group. Regarding the results of the target gene prediction, cancer and toll‑like receptor signaling were the two most significant pathways represented among the target genes, while JUN, EGFR and RAC1 were the most relevant proteins among the selected potential targets of miR‑146a‑5p. In a CAM xenograft tumor model, overexpression of miR‑146a‑5p inhibited the tumorigenesis and angiogenesis of an NSCLC cell line. miR‑146a‑5p may act as a tumor suppressor gene in NSCLC and have moderate prognostic value in lung cancer.
我们的先前研究表明，miR-146a-5p 在非小细胞肺癌（NSCLC）组织中的表达下调，这影响了 NSCLC 患者的进展和预后。因此，本研究旨在探讨 miR-146a-5p 在 NSCLC 肿瘤发生和血管生成中的功能机制。通过慢病毒转导构建 miR-146a-5p 过表达的 H460 NSCLC 细胞系后，将过表达 miR-146a-5p 的 NSCLC 细胞移植到鸡胚绒毛尿囊膜（CAM）中建立 NSCLC 鸡胚 CAM 模型。然后，测量 CAM 内肿瘤的大小，计算血管比，并通过苏木精和伊红染色观察肿瘤细胞的细胞形态、转移和炎症。使用 12 个在线软件程序预测 miR-146a-5p 的靶基因；然后使用数据库注释、可视化和综合发现 6.7 对这些基因进行基因本体富集分析和京都基因与基因组百科全书通路注释，并使用来自搜索工具检索相互作用基因/蛋白质的蛋白质-蛋白质相互作用构建蛋白质相互作用网络。与低表达组（体积 30.13±0.06mm3，血管比 16.94±0.11）相比，miR-146a-5p 过表达组（体积 6.340±0.066mm3，血管比 9.326±0.083）的异种移植肿瘤大小和血管生成情况明显受限（P<0.001）。此外，与 miR-146a-5p 低表达组的 HE 染色切片相比，观察到明显的坏死和炎症细胞浸润。关于靶基因预测的结果，癌症和 Toll 样受体信号是靶基因中最显著的两个通路，而 JUN、EGFR 和 RAC1 是 miR-146a-5p 选择的潜在靶标中最相关的蛋白质。在 CAM 异种移植肿瘤模型中，过表达 miR-146a-5p 抑制了 NSCLC 细胞系的肿瘤发生和血管生成。miR-146a-5p 可能在 NSCLC 中作为肿瘤抑制基因发挥作用，对肺癌具有中等的预后价值。
