Li F, Xiang J, Liu J, Wang X, Jiang H
Department of Tumor Radiotherapy, First Affiliated Hospital of Bengbu Medical University, Bengbu 233004, China.
Department of Respiratory and Critical Medicine, First Affiliated Hospital of Bengbu Medical University, Bengbu 233004, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2024 May 20;44(5):841-850. doi: 10.12122/j.issn.1673-4254.2024.05.05.
To explore the molecular mechanism by which FEZF1-AS1 overexpression promotes progression of nonsmall cell lung cancer (NSCLC) via the miR-130a-5p/CCND1 axis.
TCGA database was used to analyze FEZF1-AS1 expression levels in NSCLC. FEZF1-AS1 expression was detected by qRT-PCR in clinical specimens of NSCLC tissues and NSCLC cell lines, and its correlation with clinical features of the patients were analyzed. The binding sites of FEZF1-AS1 with hsa-miR-130a-5p and those of hsa-miR-130a-5p with CCND1 were predicted. CCK8 assay, clone formation assay, scratch assay, and Transwell assay were employed to examine the effects of FEZF1-AS1 knockdown and hsa-miR-130a-5p inhibitor on proliferation, invasion, and migration abilities of lung cancer cell lines. Dual luciferase assay was used to verify the binding of FEZF1-AS1 with hsa-miR-130a-5p and the binding of hsa-miR-130a-5p with CCND1. Western blotting was performed to detect the changes in CCND1 protein expression level in H1299 and H358 cells following FEZF1-AS1 knockdown and treatment with hsa-miR-130a-5p inhibitor.
FEZF1-AS1 was highly expressed in NSCLC tissues in close correlation with lymph node metastasis and also in H1299 and H358 cell lines (all < 0.05). FEZF1-AS1 knockdown obviously reduced proliferation, migration, and invasion abilities of NSCLC cells ( < 0.05). Dual luciferase assay confirmed the binding of hsa-miR-130a-5p with FEZF1-AS1 and CCND1 ( < 0.05), and hsa-miR-130a-5p inhibitor significantly inhibited proliferation, migration, and invasion of NSCLC cells ( < 0.05). FEZF1-AS1 knockdown significantly reduced CCND1 protein expression in NSCLC cells, and this effect was strongly inhibited by treatment with hsa-miR-130a-5p inhibitor ( < 0.05).
FEZF1-AS1 is highly expressed in NSCLC tissue in close correlation with lymph node metastasis to promote cancer progression through the miR-130a-5p/CCND1 axis.
探讨FEZF1-AS1过表达通过miR-130a-5p/CCND1轴促进非小细胞肺癌(NSCLC)进展的分子机制。
利用TCGA数据库分析NSCLC中FEZF1-AS1的表达水平。采用qRT-PCR检测NSCLC组织临床标本和NSCLC细胞系中FEZF1-AS1的表达,并分析其与患者临床特征的相关性。预测FEZF1-AS1与hsa-miR-130a-5p的结合位点以及hsa-miR-130a-5p与CCND1的结合位点。采用CCK8法、克隆形成法、划痕法和Transwell法检测FEZF1-AS1敲低和hsa-miR-130a-5p抑制剂对肺癌细胞系增殖、侵袭和迁移能力的影响。采用双荧光素酶报告基因检测法验证FEZF1-AS1与hsa-miR-130a-5p的结合以及hsa-miR-130a-5p与CCND1的结合。采用蛋白质免疫印迹法检测FEZF1-AS1敲低及hsa-miR-130a-5p抑制剂处理后H1299和H358细胞中CCND1蛋白表达水平的变化。
FEZF1-AS1在NSCLC组织中高表达,与淋巴结转移密切相关,在H1299和H358细胞系中也高表达(均P<0.05)。FEZF1-AS1敲低明显降低了NSCLC细胞的增殖、迁移和侵袭能力(P<0.05)。双荧光素酶报告基因检测法证实hsa-miR-130a-5p与FEZF1-AS1和CCND1结合(P<0.05),hsa-miR-130a-5p抑制剂显著抑制NSCLC细胞的增殖、迁移和侵袭(P<0.05)。FEZF1-AS1敲低显著降低NSCLC细胞中CCND1蛋白表达,hsa-miR-130a-5p抑制剂处理强烈抑制了这种作用(P<0.05)。
FEZF1-AS1在NSCLC组织中高表达,与淋巴结转移密切相关,通过miR-130a-5p/CCND1轴促进肿瘤进展。