lncRNA NEAT1通过抑制miR-let-7a激活BZW2促进喉乳头状瘤细胞的增殖和凋亡逃逸  

lncRNA NEAT1 promoting the proliferation and apoptosis escape of laryngeal papilloma cells by activating BZW2 through the inhibition of miR-let-7a

作　　者：张波[1] 王松[1] 阿依恒·曲库尔汗[1] ZHANG Bo;WANG Song;Ayiheng·Qukuerhan(Department of Otolaryngology,First Affiliated Hospital of Xinjiang Medical University,Urumqi Xinjiang 830054,China)

机构地区：[1]新疆医科大学第一附属医院耳鼻喉科,新疆乌鲁木齐830054

出　　处：《局解手术学杂志》2023年第5期382-388,共7页Journal of Regional Anatomy and Operative Surgery

基　　金：新疆维吾尔自治区区域协同创新专项项目(2020E01055)。

摘　　要：目的探讨长链非编码RNA核富集转录本1(lncRNA NEAT1)对喉乳头状瘤(LP)细胞的增殖和凋亡逃逸的调控作用和机制。方法培养人喉乳头状瘤细胞Hs840.T,将正常人口腔上皮细胞(HOECs)作为对照。将Hs840.T分为lncRNA NEAT1过表达载体(pcDNA-NEAT1)组、pcDNA-NEAT1阴性对照(pcDNA-NC)组、沉默lncRNA NEAT1的小干扰RNA载体(siNEAT1)组、siNEAT1的阴性对照(siNC)组、miR-let-7a的模拟物(mimic)组、mimic阴性对照(mimic-NC)组、miR-let-7a抑制剂(inhibitor)组、inhibitor阴性对照(inhibitor-NC)组、沉默碱性亮氨酸拉链和W2结构域2(BZW2)的小干扰RNA载体(siBZW2)组、siBZW2的阴性对照(siBZW2-NC)组、pcDNA-NEAT1联合siBZW2给药(pcDNA-NEAT1+siBZW2)组、pcDNA-NEAT1联合siBZW2-NC给药(pcDNA-NEAT1+siBZW2-NC)组。qRT-PCR检测各组中lncRNA NEAT1及其预测靶分子miR-let-7a的表达。Western blot和qRT-PCR检测BZW2的表达。双荧光素酶报告基因实验检测Hs840.T中lncRNA NEAT1和miR-let-7a以及miR-let-7a和BZW2的靶向关系。MTT实验测定Hs840.T的增殖能力。Annexin V-FITC/PI双染法检测Hs840.T的凋亡逃逸能力。结果Hs840.T中的lncRNA NEAT1和BZW2表达高于HOECs(P<0.05),而miR-let-7a表达低于HOECs(P<0.05)。双荧光素酶报告基因实验结果表明,Hs840.T中lncRNA NEAT1吸附抑制miR-let-7a并上调BZW2的表达(P<0.05),且BZW2是miR-let-7a的靶基因。与pcDNA-NC组相比,pcDNA-NEAT1组Hs840.T的增殖率增加(P<0.05),早期、晚期凋亡率均降低(P<0.05);与siNC组相比,siNEAT1组Hs840.T的增殖率降低(P<0.05),早期、晚期凋亡率均增加(P<0.05)。另外,与pcDNA-NEAT1+siBZW2-NC组相比,pcDNA-NEAT1+siBZW2组Hs840.T的增殖率降低(P<0.05),早期、晚期凋亡率均增高(P<0.05)。结论lncRNA NEAT1通过抑制miR-let-7a激活BZW2促进LP细胞的增殖和凋亡逃逸。Objective To investigate the regulatory effect of long non-coding RNA nuclear enriched abundant transcript 1(lncRNA NEAT1)on the proliferation and apoptosis escape of laryngeal papilloma(LP)cells and its mechanism.Methods Human laryngeal papilloma cells Hs840.T were cultured,and the normal human oral epithelial cells(HOECs)were used as the control.Then,the Hs840.T were divided into the lncRNA NEAT1 over-expression vector(pcDNA-NEAT1)group,pcDNA-NEAT1 negative control(pcDNA-NC)group,lncRNA NEAT1 silencing small interfering RNA vector(siNEAT1)group,siNEAT1 negative control(siNC)group,mimic of miR-let-7a(mimic)group,mimic negative control(mimic-NC)group,inhibitor of miR-let-7a(inhibitor)group,inhibitor negative control(inhibitor-NC)group,basic leucine zipper and W2 domains 2(BZW2)silencing small interfering RNA vector(siBZW2)group,siBZW2 negative control(siBZW2-NC)group,combined administration of pcDNA-NEAT1 and siBZW2(pcDNA-NEAT1+siBZW2)group,and combined administration of pcDNA-NEAT1 and siBZW2-NC(pcDNA-NEAT1+siBZW2-NC)group.qRT-PCR was used to detect the expression of lncRNA NEAT1 and its predicted target molecule miR-let-7a in each group.Western blot and qRT-PCR were used to detect the expression of BZW2.Dual-luciferase reporter gene assay was used to investigate the targeting relationship between lncRNA NEAT1 and miR-let-7a and the targeting relationship between miR-let-7a and BZW2 in Hs840.T.MTT assay was used to determine the proliferation ability of Hs840.T.Annexin V-FITC/PI double staining assay was used to determine the apoptosis escape ability of Hs840.T.Results The expression of lncRNA NEAT1 and BZW2 in Hs840.T were higher than those in HOECs(P<0.05),while the expression of miR-let-7a in Hs840.T was lower than that in HOECs(P<0.05).The results of dual-luciferase reporter gene assay showed that lncRNA NEAT1 sponged and inhibited miR-let-7a,thereby up regulating the expression of BZW2 in Hs840.T(P<0.05),and BZW2 was a target gene of miR-let-7a.Compared with the pcDNA-NC group,the pcDNA-NEAT1 group showed a

关 键 词：喉乳头状瘤 长链非编码RNA核富集转录本1 miR-let-7a 碱性亮氨酸拉链和W2结构域2 细胞凋亡 细胞增殖 

分 类 号：R739.65[医药卫生—肿瘤]