miR-101-5p通过靶向ATAD2抑制肺鳞癌细胞的侵袭  

MiR-101-5p inhibits the proliferation and invasion of lung squamous cell carcinoma cells by targeting ATAD2

作　　者：刘浩杰 王德才 李树斌 LIU Haojie;WANG Decai;LI Shubin(Department of Cardiothoracic Surgery,The Fourth Clinical College of Xinxiang Medical College,Xinxiang Central Hospital,Xinxiang 453000,China)

机构地区：[1]新乡市中心医院/新乡医学院第四临床学院心胸外科,河南新乡453000

出　　处：《西安交通大学学报（医学版）》2024年第2期228-236,共9页Journal of Xi’an Jiaotong University（Medical Sciences）

摘　　要：目的 探讨微小RNA-101-5p(miR-101-5p)影响肺鳞癌(lung squamous cell carcinoma,LUSC)细胞增殖、侵袭的分子机制。方法 从癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库下载肺鳞癌TCGA-LUSC的miRNA成熟体表达数据和总RNA的测序数据,鉴定差异表达基因;分析富集于ATAD2的信号通路。培养LUSC细胞,qRT-PCR检测细胞中miR-101-5p和ATAD2的mRNA表达,MTT实验、克隆形成实验和侵袭实验检测miR-101-5p对LUSC细胞增殖和侵袭的影响,流式细胞术检测ATAD2对LUSC细胞周期的影响,Western blotting检测ATAD2蛋白的表达。双荧光素酶实验验证miR-101-5p能否与ATAD2靶向结合,最后通过LUSC细胞共转染oe-ATAD2和miR-101-5p模拟物(mimic),检测细胞增殖、克隆、侵袭能力的变化,进一步探究miR-101-5p能否通过ATAD2调节LUSC细胞的生物学功能。结果 miR-101-5p在LUSC组织及培养LUSC细胞中显著下调。过表达miR-101-5p的LUSC增殖和侵袭能力显著抑制。其下游调控靶基因ATAD2在LUSC中显著上调,且miR-101-5p和ATAD2表达呈负相关,单基因富集分析(GSEA)结果显示ATAD2显著富集于细胞周期信号通路。双荧光素酶实验结果显示,miR-101-5p靶向结合ATAD2,并通过MTT和侵袭实验发现miR-101-5p通过靶向结合ATAD2调控LUSC细胞的增殖和侵袭。结论 miR-101-5p在LUSC中低表达,miR-101-5p通过靶向抑制ATAD2降低LUSC细胞的增殖、侵袭。Objective To investigate the molecular mechanism of microRNA-101-5p(miR-101-5p) affecting the proliferation and invasion of lung squamous cell carcinoma(LUSC) cells.Methods We downloaded the miRNA mature expression data and total RNA sequencing data of TCGA-LUSC from TCGA database to identify differentially expressed genes.The signal pathway enriched in ATAD2 was analyzed.The mRNA expressions of miR-101-5p and ATAD2 in the LUSC cells were detected by qRT-PCR.The effects of miR-101-5p on the proliferation and invasion of LUSC cells were detected by MTT assay,cloning assay,and invasion assay.The effects of ATAD2 on the cell cycle of LUSC cells were detected by flow cytometry.Western blotting was used to detect the expression of ATAD2 protein.Double luciferase experiment was used to verify whether miR-101-5p could bind to ATAD2 target.Finally,we detected the changes in the proliferation,cloning and invasion ability of LUSC cells by co-transfection with oe-ATAD2 and miR-101-5p mimic,and further explored whether miR-101-5p could regulate the biological function of LUSC cells through ATAD2.Results The miR-101-5p was significantly downregulated in LUSC tissues and cells.Overexpression of miR-101-5p could significantly inhibit the proliferation and invasion of LUSC cells.ATAD2,its downstream regulatory target gene,was significantly upregulated in LUSC,and miR-101-5p and ATAD2 expressions were inversely correlated.GSEA enrichment results showed that ATAD2 was significantly enriched in the cell cycle signal pathway.The double luciferase experiment proved that miR-101-5p targeted ATAD2,and the recovery experiment showed that miR-101-5p regulated the proliferation and invasion of LUSC cells by targeting ATAD2.Conclusion In this study,we found that miR-101-5p had low expression in LUSC,and that miR-101-5p decreased the proliferation and invasion of LUSC cells by targeted inhibition of ATAD2.

关 键 词：微小RNA-101-5p(miR-101-5p) ATP酶家族AAA域(ATAD2) 肺鳞癌(LUSC) 增殖 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]