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中国癌症防治杂志

中国癌症防治杂志 ›› 2017, Vol. 9 ›› Issue (4): 277-282.doi: 10.3969/j.issn.1674-5671.2017.04.07

• 鼻咽癌专栏 • 上一篇    下一篇

下调叉头框转录因子3a促进鼻咽癌侵袭转移

  

  1. 华中科技大学同济医学院附属同济医院肿瘤科
  • 出版日期:2017-08-25 发布日期:2017-09-12
  • 通讯作者: 胡国清。E-mail：gqhu@tjh.tjmu.edu.cn
  • 基金资助:

    国家自然科学基金资助项目（81272491，81572960）

Silencing Forkhead Box Transcription Factor（FOXO3a） promotes tumor invasion and migration in nasopharyngeal carcinoma

  1. Department of Oncology，Tongji Hospital，Tongji Medical CollegeAffiliated，Huazhong University of Science and Technology
  • Online:2017-08-25 Published:2017-09-12
  • Contact: Hu Guoqing. Email：gqhu@tjh.tjmu.edu.cn

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摘要：

目的 探讨下调叉头框转录因子3a（FOXO3a）对促进鼻咽癌侵袭转移的影响。方法 慢病毒shRNA和空载病毒液Mock shRNA按复感染指数（multiplicity of infection，MOI）为50转染鼻咽癌CNE2和HNE1细胞，构建低表达FOXO3a的稳定转染细胞。采用Western blot、Real-time PCR检测其转染效率。划痕实验和Transwell 实验检测下调FOXO3a对诱导鼻咽癌细胞侵袭转移的影响。Western blot检测上皮间质转化（epithelial-mesenchymal transition，EMT）相关蛋白 E-cadherin、Vimentin、N-cadherin、Twist和Snail的表达，免疫荧光检测EMT相关蛋白定位。Western blot和Real-time PCR检测基质金属蛋白酶MMP2和MMP9的表达。结果 成功构建低表达FOXO3a稳定转染鼻咽癌CNE2和HNE1细胞。划痕实验和Transwell 实验结果显示，与空载体转染鼻咽癌细胞比较，低表达FOXO3a稳定转染鼻咽癌细胞的侵袭和转移能力增强，EMT相关蛋白E-cadherin表达下调，而Vimentin、N-cadherin、Twist和Snail表达显著升高，MMP2和MMP9表达亦升高。结论 下调FOXO3a可促进鼻咽癌的侵袭转移。

关键词: 鼻咽肿瘤, 叉头框转录因子3a, 侵袭, 转移

Abstract:

Objective To investigate whether Silencing Forkhead Box Transcription Factor（FOXO3a） can promote tumor invasion and migration in nasopharyngeal carcinoma（NPC）. Methods The NPC cell lines CNE2 and HNE1 were transfected with lentiviruses encoding short hairpin RNA（shRNA） targeting FOXO3a or encoding mock shRNA，and transfection efficiency was checked using Western blotting and real-time PCR. Scratch and transwell tests were performed to determine whether silencing FOXO3a induced invasion and migration of NPC cells. Western blotting was performed to identify changes in levels of the epithelial-mesenchymal transition markers E-cadherin，Vimentin，N-cadherin，Twist and Snail. Localization of these marker proteins was analyzed using immunofluorescence. Western blotting and real-time PCR were also used to measure expression of matrix metalloproteinases（MMPs） 2 and 9. Results Stably transfected cell lines were constructed in which FOXO3a was silenced. Silencing FOXO3a promoted tumor invasion and migration，based on the scratch and transwell tests. Levels of the epithelial marker E-cadherin were reduced. In contrast，levels of the mesenchymal markers vimentin and N-cadherin were increased，as were levels of MMP2 and MMP9. Conclusion Silencing FOXO3a may promote tumor invasion and migration in NPC.

Key words: Nasopharyngeal neoplasm, FOXO3a, Invasion, Migration