外泌体介导的miR-577在幼儿复发性喉乳头状瘤中的表达及其对细胞增殖和侵袭的影响  

Expression of Exosome Mediated miR-577 in Children with Recurrent Laryngeal Papilloma and Its Effect on Proliferation and Invision of Laryngeal Papilloma Cells

作　　者：胡彬雅[1] 李赟[1] 敬云龙[1] 赵斯君[1] Hu Binya;Li Yun;Jing Yunlong;Zhao Sijun(Hunan Children's Hospital Otorhinolaryngology,Head and Neck Surgery,Changsha,410007,China)

机构地区：[1]湖南省儿童医院耳鼻咽喉头颈外科,长沙410007

出　　处：《听力学及言语疾病杂志》2022年第1期5-11,共7页Journal of Audiology and Speech Pathology

基　　金：湖南省卫生计生委科研基金课题(B2011-093)。

摘　　要：目的探讨miR-577在幼儿复发性喉乳头状瘤中的表达及其对细胞增殖、侵袭的影响和潜在分子机制。方法收集2016年6月至2019年12月确诊的30例幼儿喉乳头状瘤组织样本及其相匹配且远离病灶经病理检查证实的正常喉组织样本,并选取人正常喉上皮细胞株和喉乳头状瘤细胞(分为miR-577过表达组、阴性对照组及空白对照组)进行研究。采用实时荧光定量PCR(qRT-PCR)分别检测30例喉乳头状瘤组织、配对正常喉组织、正常喉上皮细胞和喉乳头状瘤细胞中miR-577相对表达;通过Lipofectamine^(TM)3000脂质体转染试剂盒将miR-577 mimic模拟物转染至喉乳头状瘤细胞构建miR-577过表达细胞系,采用CCK-8法、Tanswell实验分别检测miR-577对喉乳头状瘤细胞增殖、侵袭的影响;采用Western blot检测miR-577对喉乳头状瘤细胞侵袭相关蛋白(MMP-1、MMP-9)表达及Notch通路相关蛋白(Notch、DSL)表达的影响。结果幼儿喉乳头状瘤组织中miR-577相对表达量(0.134±0.026)显著低于正常喉组织(0.823±0.056)(P<0.001);喉乳头状瘤细胞中miR-577相对表达量(0.256±0.026)显著低于正常喉上皮细胞(0.962±0.072)(P<0.001);与阴性对照组相比,转染miR-577 mimic后喉乳头状瘤细胞增殖能力(1.248±0.103 vs 0.902±0.094)和细胞侵袭率(71.016±5.869 vs 42.867±4.795)明显降低(P<0.05);侵袭相关蛋白MMP-1(1.011±0.124 vs 0.417±0.056)、MMP-9(1.012±0.155 vs 0.502±0.058)表达抑制(P<0.01)。喉乳头状瘤细胞中Notch、DSL表达显著上调(P<0.05);转染miR-577 mimic后喉乳头状瘤细胞中Notch通路相关蛋白Notch、DSL表达明显抑制(P<0.05)。共转染miR-577 mimic和pcDNA3.1-Notch后喉乳头状瘤细胞增殖能力和细胞侵袭率较单纯转染miR-577 mimic组明显增加(P<0.05);共转染pcDNA3.1-Notch逆转了miR-577 mimic对喉乳头状瘤细胞增殖、侵袭能力的抑制作用。结论幼儿喉乳头状瘤组织及喉乳头状瘤细胞中miR-577显著低表达,过表达miR-577显著Objective To investigate the expression of miR-577 in recurrent laryngeal papilloma in children and its effect on proliferation and invasion of laryngeal papilloma cells and its potential molecular mechanism.Methods Tissue samples from 30 cases of laryngeal papilloma confirmed in our hospital from June 2016 to December 2019 were collected and matched with normal laryngeal tissue samples.Human normal laryngeal epithelial cell lines and laryngeal papilloma cells were selected for study.Quantitative real-time PCR(qRT-PCR)was used to detect the relative expression of miR-577 in 30 cases of laryngeal papilloma tissues and paired normal laryngeal tissues,as well as in normal laryngeal epithelial cells and laryngeal papilloma cells.MiR-577 overexpressed cell lines were constructed by transfecting miR-577 mimic into laryngeal papilloma cells using LipofectamineTM3000 liposome transfection kit.The effects of miR-577 on proliferation and invasion of laryngeal papilloma cells were detected by CCK-8 method and Tanswell experiment,respectively.Western blot was used to detect the effect of miR-577 on the expression of invasion related proteins(MMP-1 and MMP-9)and Notch pathway related proteins(Notch and DSL)in laryngeal papilloma cells.Results The relative expression of miR-577 in the laryngeal papilloma tissues of children(0.134±0.026 vs 0.823±0.056)was significantly lower than that in the normal laryngeal tissues(P<0.001).The relative expression of miR-577 in laryngeal papilloma cells(0.256±0.026 vs 0.962±0.072)was significantly lower than that in normal laryngeal epithelial cells(P<0.001).Compared with the NC mimic group,proliferation ability of laryngeal papilloma cells after transfection with miR-577 mimic(1.248±0.103 vs 0.902±0.094)and cell invasion rate(71.016±5.869 vs 42.867±4.795)were significantly reduced(P<0.05).The expression of invasion-related proteins MMP-1(1.011±0.124 vs 0.417±0.056)and MMP-9(1.012±0.155 vs 0.502±0.058)was inhibited(P<0.01).The expression of Notch and DSL in laryngeal papilloma cell

关 键 词：miR-577 喉乳头状瘤 增殖 侵袭 NOTCH信号通路 

分 类 号：R764.43[医药卫生—耳鼻咽喉科]