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.
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.
李菲凡;向俊馨;刘佳慧;王效静;江浩
蚌埠医科大学第一附属医院 肿瘤放疗科,安徽 蚌埠 233004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||綦江区人民医院呼吸与危重症医学科,重庆 401420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||呼吸系病临床基础安徽省重点实验室,安徽 蚌埠 233004
非小细胞肺癌;长链非编码RNA;FEZF1-AS1;miR-130a-5p;CCND1
non-small cell lung cancer;lncRNA;FEZF1-AS1;miR-130a-5p;CCND1
《南方医科大学学报》 2024 (005)
841-850 / 10
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) Supported by National Natural Science Foundation of China(82373329).
10.12122/j.issn.1673-4254.2024.05.05
评论