lncRNA PART1通过miR-204-3p/IGFBP2轴促进非小细胞肺癌的实验研究  

Experimental study of lncRNA PART1 promoting non-small cell lung cancer through miR-204-3p/IGFBP2 axis

作　　者：陈建广 王芳 王秀云 CHEN Jian-guang;WANG Fang;WANG Xiu-yun(Department of Thoracic Surgery,Dongying People′s Hospital,Dongying,Shandong 257091,China)

机构地区：[1]东营市人民医院胸外科,山东东营257091

出　　处：《临床肺科杂志》2022年第5期736-743,共8页Journal of Clinical Pulmonary Medicine

摘　　要：目的通过体外细胞研究和动物研究,探讨长链非编码RNA前列腺雄激素调节转录本1(PART1)通过miR-204-3p/胰岛素样生长因子结合蛋白2(IGFBP-2)轴对非小细胞肺癌(NSCLC)的影响。方法用RT-PCR法检测NSCLC细胞系A549和PC9,肺腺癌细胞系H1299、H1650和H1975,人正常支气管上皮细胞系(HBE)中PART1的相对表达量。对A549细胞转染shRNA-NC或shRNA-PART1,分析敲降PART1对癌细胞增殖、侵袭、迁移和凋亡的影响,将10只BALB/c裸鼠随机分为sh-NC组和sh-PART1组,分析PART1对肿瘤增殖的影响。用双荧光素酶报告基因验证PART1、miR-204-3p和IGFBP2的靶向作用关系。对A549细胞分别仅转染shRNA-PART1、同时转染PART1-shRNA+miR-204-3p inhibitor、同时转染PART1 shRNA+pcDNA-IGFBP2,分析PART1通过miR-204-3p/IGFBP2轴,对细胞增殖、侵袭、迁移和凋亡的影响。克隆形成实验和CCK-8法检测细胞增殖能力,Transwell小室实验检测细胞侵袭能力,体外损伤愈合实验检测细胞迁移。结果与HBE细胞比较,A549、H1299、H1650、H1975和PC9细胞中PART1相对表达量明显较高(P<0.05)。敲降PART1可以明显抑制A549细胞的增殖、侵袭和迁移能力(P<0.05)。在动物实验中,抑制PART1的表达可以明显降低肿瘤的体积和重量(P<0.05)。双荧光素酶报告基因检测证实,PART1靶向作用于miR-204-3p并下调其表达,miR-204-3p可负调控IGFBP-2的表达。敲降PART1表达可以明显抑制A549细胞中IGFBP-2 mRNA和蛋白的表达水平(P<0.05)。同时,转染PART1-shRNA+miR-204-3p inhibitor或者PART1 shRNA+pcDNA-IGFBP2后,IGFBP-2 mRNA和蛋白的表达水平较仅转染PART1-shRNA明显上调(P<0.05),细胞增殖、侵袭和迁移能力也明显升高(P<0.05)。结论通过体外细胞研究和动物研究,本研究证实PART1可通过调控miR-204-3p/IGFBP2轴促进NSCLC的进展。Objective To explore the effect of long-chain non-coding RNA prostate androgen-regulated transcript 1(PART1)on non-small cell lung cancer(NSCLC)via the miR-204-3p/insulin-like growth factor binding protein 2(IGFBP-2)axis by in vitro and animal experiments.Methods RT-PCR was used to detect the relative expression of PART1 in NSCLC cell lines A549 and PC9,lung adenocarcinoma cell lines H1299,H1650,and H1975,and human normal bronchial epithelial cell line(HBE).A549 cellswere transfected with shRNA-NC or shRNA-PART1 to analyze the effect of knockdown PART1 on cancer cell proliferation,invasion,migration,and apoptosis.10 BALB/c nude mice were randomly divided into sh-NC group and sh-PART1 group to analyze the effect of PART1 on tumor proliferation.The dual-luciferase reporter gene was used to verify the targeting relationship between PART1,miR-204-3p,and IGFBP2.A549 cells were transfected with only shRNA-PART1,PART1-shRNA+miR-204-3p inhibitor,and PART1 shRNA+pcDNA-IGFBP2,respectively,to analyze the effect of PART1on cell proliferation,invasion,migration,and apoptosis through the miR-204-3p/IGFBP2 axis.Clone formation experiment and CCK-8 method were used to detect cell proliferation ability.A Transwell chamber experiment was used to detect cell invasion ability,and a cell scratch experiment was used to detect cell migration.Results Compared with HBE cells,the relative expression of PART1 in A549,H1299,H1650,H1975,and PC9 cells was significantly higher(P<0.05).Knockdown of PART1 could significantly inhibit the proliferation,invasion,and migration of A549 cells(all P<0.05).In animal experiments,inhibiting the expression of PART1 could significantly reduce tumor volume and weight(P<0.05).The dual-luciferase reporter gene assay confirmed that PART1 targets miR-204-3p and down-regulates its expression,and miR-204-3p could negatively regulate the expression of IGFBP-2.Knockdown of PART1 expression could significantly inhibit the expression levels of IGFBP-2 mRNA and protein in A549 cells(all P<0.05).Meanwhile,after transfec

关 键 词：长链非编码RNA 前列腺雄激素调节转录本1 微小RNA 胰岛素样生长因子结合蛋白2 

分 类 号：R734.2[医药卫生—肿瘤]