Chen Xiaoying, Jiang Ruilai, Huang Xiaocheng, Chen Ling, Hu Xiaogang, Wei Yanbin
Department of Respiratory and Critical Care Medicine, The Second Peoples' Hospital of Lishui, Lishui, China.
Cell Biochem Biophys. 2025 Mar;83(1):415-427. doi: 10.1007/s12013-024-01472-w. Epub 2024 Aug 8.
Non-small cell lung cancer (NSCLC) is the most common malignancy worldwide, with a high death rate. Long noncoding RNA (LncRNA) NKX2-1 antisense RNA 1 (NKX2-1-AS1) has been reported to be an oncogene in lung tumorigenesis. However, the precise mechanism of NKX2-1-AS1 underlying NSCLC progression is blurry. The intention of our research was to probe the potential mechanism of NKX2-1-AS1 underlying NSCLC. NKX2-1-AS1 expression and relevant downstream gene expression were measured using RT-qPCR. Cell proliferation and apoptosis were determined by MTT assay, EdU assay along with flow cytometry analysis. Cell migratory and invasive abilities were inspected by transwell assay. Western blot and immunofluorescence staining were utilized to assess the levels of epithelial-mesenchymal transition (EMT)-related proteins. RNA pull-down together with luciferase reporter assays were performed to verify the interaction between NKX2-1-AS1 and its downstream RNAs. Xenograft tumor-bearing mouse models were built to analyze tumor growth in vivo. The results suggested that NKX2-1-AS1 was upregulated in NSCLC patient tissues and cell lines. NKX2-1-AS1 deficiency suppressed cell proliferation, migration, invasion and EMT while elevated apoptosis. NKX2-1-AS1 bound to miR-589-5p, and NME/NM23 nucleoside diphosphate kinase 1 (NME1) was targeted by miR-589-5p in NSCLC cells. Additionally, NKX2-1-AS1 accelerated the progression of NSCLC by regulating miR-589-5p/NME1 axis. NKX2-1-AS1 knockdown repressed tumor growth in vivo. In conclusion, NKX2-1-AS1 accelerated the NSCLC progression through interacting with miR-589-5p for NME1 upregulation, which may provide clues for NSCLC targeting therapy.
非小细胞肺癌（NSCLC）是全球最常见的恶性肿瘤，死亡率很高。据报道，长链非编码RNA（LncRNA）NKX2-1反义RNA 1（NKX2-1-AS1）在肺癌发生过程中是一种致癌基因。然而，NKX2-1-AS1促进NSCLC进展的确切机制尚不清楚。我们研究的目的是探究NKX2-1-AS1促进NSCLC的潜在机制。使用RT-qPCR检测NKX2-1-AS1的表达及相关下游基因的表达。通过MTT法、EdU法以及流式细胞术分析来测定细胞增殖和凋亡情况。通过Transwell实验检测细胞迁移和侵袭能力。利用蛋白质免疫印迹法和免疫荧光染色法评估上皮-间质转化（EMT）相关蛋白的水平。进行RNA下拉实验和荧光素酶报告基因检测以验证NKX2-1-AS1与其下游RNA之间的相互作用。构建异种移植荷瘤小鼠模型以分析体内肿瘤生长情况。结果表明，NKX2-1-AS1在NSCLC患者组织和细胞系中表达上调。NKX2-1-AS1缺失可抑制细胞增殖、迁移、侵袭和EMT，同时增加细胞凋亡。NKX2-1-AS1与miR-589-5p结合，在NSCLC细胞中，NME/NM23核苷二磷酸激酶1（NME1）是miR-589-5p的靶标。此外，NKX2-1-AS1通过调节miR-589-5p/NME1轴加速NSCLC的进展。敲低NKX2-1-AS1可抑制体内肿瘤生长。总之，NKX2-1-AS1通过与miR-589-5p相互作用上调NME1从而加速NSCLC进展，这可能为NSCLC的靶向治疗提供线索。
