南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (5): 841-850.doi: 10.12122/j.issn.1673-4254.2024.05.05

• 基础研究 • 上一篇    下一篇

LncRNA FEZF1-AS1通过miR-130a-5p/CCND1轴促进非小细胞肺癌发展的分子机制研究

李菲凡1(), 向俊馨2,3(), 刘佳慧2, 王效静2,4, 江浩1()   

  1. 1.蚌埠医科大学第一附属医院,肿瘤放疗科,安徽 蚌埠 233004
    2.蚌埠医科大学第一附属医院,呼吸与危重症医学科,安徽 蚌埠 233004
    3.綦江区人民医院呼吸与危重症医学科,重庆 401420
    4.呼吸系病临床基础安徽省重点实验室,安徽 蚌埠 233004
  • 收稿日期:2023-12-27 出版日期:2024-05-20 发布日期:2024-06-04
  • 通讯作者: 江浩 E-mail:945712119@qq.com;326089784@qq.com;Jianghao1223@163.com
  • 作者简介:李菲凡,在读硕士研究生,医师,E-mail: 945712119@qq.com
    向俊馨,硕士,医师,E-mail: 326089784@qq.com
    第一联系人::李菲凡、向俊馨共同为第一作者
  • 基金资助:
    国家自然科学基金(82373329);安徽省教育厅自然科学研究重点项目(KJ2018A0997)

Overexpression of lncRNA FEZF1-AS1 promotes progression of non-small cell lung cancer via the miR-130a-5p/CCND1 axis

Feifan LI1(), Junxin XIANG2,3(), Jiahui LIU2, Xiaojing WANG2,4, Hao JIANG1()   

  1. 1.Department of Tumor Radiotherapy, First Affiliated Hospital of Bengbu Medical University, Bengbu 233004, China
    2.Department of Respiratory and Critical Medicine, First Affiliated Hospital of Bengbu Medical University, Bengbu 233004, China
    3.Department of Respiratory and Critical Medicine, Qijiang District People's Hospital, Chongqing 401420, China
    4.Clinical Basis of Respiratory Diseases Key Laboratory of Anhui Province, Bengbu 233004, China
  • Received:2023-12-27 Online:2024-05-20 Published:2024-06-04
  • Contact: Hao JIANG E-mail:945712119@qq.com;326089784@qq.com;Jianghao1223@163.com
  • Supported by:
    National Natural Science Foundation of China(82373329)

摘要:

目的 探讨FEZF1-AS1通过miR-130a-5p/CCND1轴促进非小细胞肺癌(NSCLC)发展的分子机制。 方法 利用TCGA数据库分析FEZF1-AS1在NSCLC中的表达,qRT-PCR检测其在NSCLC癌组织与癌旁组织以及NSCLC细胞系中的表达,并分析其与临床特征的关系。利用数据库预测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;应用CCK8、克隆形成实验、划痕实验、Transwell实验检测FEZF1-AS1、hsa-miR-130a-5p对肺癌细胞系增殖、侵袭、迁移能力的影响;双荧光素酶报告实验验证FEZF1-AS1与hsa-miR-130a-5p以及hsa-miR-130a-5p与CCND1存在结合;将H1299、H358分别分为si-NC组、si-FEZF1-AS1组、si-FEZF1-AS1+NC inhibitor组、si-FEZF1-AS1+hsa-miR-130a-5p inhibitor组,应用Western blot检测CCND1的蛋白表达水平,明确FEZF1-AS1/hsa-miR-130a-5p是否通过ceRNA机制调控了CCND1的表达。 结果 FEZF1-AS1在NSCLC中呈高表达,且在NSCLC癌组织中的表达量明显增高(P<0.05),高表达与NSCLC的淋巴结转移有关,在H1299、H358细胞系中的表达量也显著增高(P<0.05);敲减FEZF1-AS1降低了NSCLC细胞的增殖、迁移、侵袭能力(P<0.05);hsa-miR-130a-5p与FEZF1-AS1、CCND1之间存在结合(P<0.05);hsa-miR-130a-5p影响NSCLC细胞的增殖、迁移、侵袭能力(P<0.05);si-FEZF1-AS1降低了CCND1蛋白表达,hsa-miR-130a-5p inhibitor逆转了si-FEZF1-AS对CCND1的敲减作用(P<0.05)。 结论 FEZF1-AS1在NSCLC组织中高表达,且与淋巴结转移有关,促进了NSCLC细胞的增殖、迁移、侵袭,并通过miR-130a-5p/CCND1轴促进了非小细胞肺癌的发展。

关键词: 非小细胞肺癌, 长链非编码RNA, FEZF1-AS1, miR-130a-5p, CCND1

Abstract:

Objective To explore the molecular mechanism by which FEZF1-AS1 overexpression promotes progression of non-small cell lung cancer (NSCLC) via the miR-130a-5p/CCND1 axis. Methods 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. Results FEZF1-AS1 was highly expressed in NSCLC tissues in close correlation with lymph node metastasis and also in H1299 and H358 cell lines (all P<0.05). FEZF1-AS1 knockdown obviously reduced proliferation, migration, and invasion abilities of NSCLC cells (P<0.05). Dual luciferase assay confirmed the binding of hsa-miR-130a-5p with FEZF1-AS1 and CCND1 (P<0.05), and hsa-miR-130a-5p inhibitor significantly inhibited proliferation, migration, and invasion of NSCLC cells (P<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 (P<0.05). Conclusion 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.

Key words: non-small cell lung cancer, lncRNA, FEZF1-AS1, miR-130a-5p, CCND1