实用医学杂志 ›› 2021, Vol. 37 ›› Issue (3): 314-318.doi: 10.3969/j.issn.1006⁃5725.2021.03.008

• 基础研究 • 上一篇    下一篇

LncRNA HOTAIR通过调控miR⁃30d影响鼻咽癌细胞的侵袭和迁移

陈曦,朱悦莹,施典羽,颜帅,汤国栋,邹宇
  

  1. 1 广东省妇幼保健院耳鼻咽喉科(广州 511400);2 深圳市龙华区人民医院耳鼻咽喉科(广东深圳 518109)
  • 出版日期:2021-02-10 发布日期:2021-02-10
  • 通讯作者: 邹宇 E⁃mail:zouyudiyouxiang@sina.com
  • 基金资助:
    深圳市科技计划项目(编号:JCYJ20170307141944428)

Influence of LncRNA HOTAIR on invasion and migration of nasopharyngeal carcinoma cells by regulating miR⁃30d

CHEN Xi,ZHU Yueying,SHI Dianyu,YAN Shuai,TANG Guodong,ZOU Yu   

  1. Department of Oto⁃rhinolaryngology,Guangdong Women′s and Children′s Hospital,Guangzhou 511400,China;*Department of Oto⁃rhinolaryngology,Longhua District People′s Hospital,Shenzhen 518109,China
  • Online:2021-02-10 Published:2021-02-10
  • Contact: ZOU Yu E⁃mail:zouyudiyouxiang@sina.com

摘要: 目的 探讨长链非编码RNA(LncRNA)HOX 转录本反义RNA(HOTAIR)通过调控靶向小分子 RNA⁃30d(miR⁃30d)影响鼻咽癌细胞侵袭和迁移的机制。方法 在鼻咽癌 CNE⁃2 细胞中转染 HOTAIR siRNA,使用实时定量聚合酶链反应(RT⁃qPCR)测定转染效果,Transwell 法测定肿瘤细胞侵袭及迁移能力,蛋白质免疫印迹(Western blot)检测细胞中上皮型钙黏蛋白(E⁃cadherin)、波形蛋白(vimentin)和葡萄糖 调节蛋白 78(GRP78)的表达;生物信息学软件分析 HOTAIR  miR⁃30d 区域结合位点,利用双荧光素酶报 告系统确定两者结合关系;用 RT⁃qPCR 方法检测下调 HOTAIR 后鼻咽癌细胞中 miR⁃30d 表达的变化。将 HOTAIR siRNA  miR⁃30d inhibitor 共转染至CNE⁃2细胞中,用上述方法分析细胞侵袭、迁移及E⁃cadherin vimentin  GRP78 蛋白表达的变化。结果 敲减 HOTAIR 表达后,CNE⁃2 细胞的侵袭和迁移能力下降,细胞中 vimentinGRP78 蛋白水平降低,E⁃cadherin蛋白水平升高;HOTAIR 靶向调控 miR⁃30d 的表达,敲减 HOTAIR 的表达可以提高CNE⁃2细胞中miR⁃30d的水平,miR⁃30d inhibitor可以明显逆转敲减 HOTAIR  CNE⁃2细胞侵袭、迁移能力以及E⁃cadherin、vimentin和GRP78蛋白表达的影响。 HOTAIR通过负调控 miR⁃30d影响GRP78和上皮细胞⁃间充质转化(EMT

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Abstract:

Objective To investigate the mechanism of long⁃chain non⁃coding RNA(LncRNA)HOX transcript antisense RNA(HOTAIR)targeting microRNA ⁃30d(miR ⁃30d)on invasion and migration of nasopharyngeal carcinoma cells. Methods HOTAIR siRNA was transfected into CNE ⁃2 cells of nasopharyngeal carcinoma. Real ⁃ time quantitative polymerase chain reaction(RT⁃qPCR)was performed to measure the transfection effect. Transwell assay was carried out to measure the invasion and migration abilities of tumor cell.Western blot was used to detect the expression of epithelial cadherin,vimentin and glucose regulatory protein78(GRP78)in the cells. Bioinformatics software was used to analyze the binding sites of HOTAIR and miR⁃30d. The binding relationship was determined by double luciferase reporting system.RT⁃qPCR was used to detect the change of miR⁃30d expression after HOTAIR was down⁃regulated in nasopharyngeal carcinoma cells. HOTAIR siRNA and miR⁃30d inhibitor were co⁃transfected into CNE⁃2 cells.The changes of cell invasion and migration were analyzed by the above methods,as well as the expression of E⁃cadherin,vimentin and GRP78 proteins. Results After knocking down the expression of HOTAIR,the invasion and migration abilities of CNE⁃2 cells were decreased;the levels of vimentin and GRP78 were decreased but the level of E⁃cadherin was increased.HOTAIR regulated specifically the expression of miR⁃30d. The knock⁃down of HOTAIR expression could increase the level of miR⁃30d in CNE⁃2 cells.The miR⁃30d inhibitor significantly reversed the effects of HOTAIRsiRNA on invasion and migration abilities of CNE⁃2 cells and the expression of E⁃cadherin,vimentin and GRP78 proteins. Conclusion HOTAIR affects the expression of GRP78 and EMT⁃related proteins by negatively regulating miR⁃30d,

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