MicroRNA-21对儿童中耳胆脂瘤细胞增殖和凋亡的调控作用  

Effect of MicroRNA-21 in Proliferation and Apoptosis in Pediatric Middle Ear Cholesteatoma Keratinocyts

作　　者：陈晓华[1] 秦兆冰[1] 

机构地区：[1]郑州大学第一附属医院耳科,郑州450052

出　　处：《听力学及言语疾病杂志》2017年第4期385-388,共4页Journal of Audiology and Speech Pathology

基　　金：河南省教育厅科技攻关项目(13A320649)

摘　　要：目的探讨MicroRNA-21(miR-21)及其靶基因第10号染色体缺失的磷酸酶及张力蛋白同源的基因(PTEN)和程序性细胞死亡因子4(PDCD4)蛋白在儿童中耳胆脂瘤细胞增殖和凋亡中的调控作用。方法选取2013年8月至2015年3月儿童中耳胆脂瘤患者手术标本23例,提取并培养其中耳胆脂瘤角质细胞,分为实验组、阴性对照组和空白对照组,实验组以介导miR-21抑制体的慢病毒载体转染中耳胆脂瘤角质细胞,阴性对照组转染空慢病毒载体,空白对照组不进行处理,利用实时定量PCR(qRT-PCR)和Western blot检测各组细胞中miR-21和PTEN基因、PDCD4蛋白的表达,利用EdU和TUNEL法检测各组细胞的增殖和凋亡情况。结果实验组胆脂瘤角质细胞中miR-21表达显著低于空白对照组和阴性对照组(P<0.05),PTEN基因和PDCD4蛋白的表达明显高于空白对照组和阴性对照组(P<0.05),EdU阳性细胞显著低于阴性对照组和空白对照组(P<0.05),TUNEL阳性细胞显著高于阴性对照组和空白对照组(P<0.05)。结论 MiR-21可负性调控其靶基因PTEN和PDCD4的表达,miR-21抑制体可有效抑制儿童胆脂瘤角质细胞的增殖活性,促进细胞的凋亡。Objective This study aims to investigate the post-transcriptional regulatory effects that control proliferation,apoptosis,and invasion in pediatriccholesteatoma keratinocytes.In particular,the potential role of miR-21 was focused on in this study.Methods A total of 23 pediatric cholesteatoma tissues were processed for cell culture.Pediatriccholesteatoma keratinocytes were transfected with miR-21 inhibitors,or negative control miRNAs.RT-PCR was used to assess the expression levels of miR-21.EdU incorporation assay and TUNEL staining were used to assess the proliferation and apoptosis of pediatric cholesteatomakeratinocytes,respectively.Results MiRNA-21 was downregulated when pediatric cholesteatoma keratinocytes were transfected with miR-21 inhibitors.Furthermore,the number of proliferative EdU＋cells decreased in cholesteatoma keratinocytes transfected with miR-21inhibitors;and the number of TUNEL-positive cells also increased in cells transfected with miR-21 inhibitors,compared with cells transfected control miRNA.Conclusion MiRNA-21 promotes the proliferation and inhibits apoptosis of pediatric cholesteatoma keratinocytes.

关 键 词：MICRORNA-21 儿童 中耳胆脂瘤 PTEN基因 PDCD4蛋白 

分 类 号：R764.21[医药卫生—耳鼻咽喉科]