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sh RNA和空载病毒液Mock sh RNA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。
Objective To investigate whether Silencing Forkhead Box and migration in nasopharyngeal carcinoma （NPC）. Methods The NPC encoding short hairpin RNA （shRNA） targeting FOXO3a or encoding Transcription Factor （FOXO3a） can promote tumor invasion cell lines CNE2 and HNE1 were transfected with lcntiviruscs 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-eadherin,Vimentin,N-eadherin,Twist and Snail. Localization of these marker proteins was analyzed using 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.
出处
《中国癌症防治杂志》
CAS
2017年第4期277-282,共6页
CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT
基金
国家自然科学基金资助项目(81272491
81572960)
