摘要
目的构建ebv-miR-BART7慢病毒表达载体,建立其稳定表达的鼻咽癌细胞株CNE1-pLVTHM/BART7。方法构建慢病毒表达质粒pLVTHM/BART7。用293FT细胞包装后产生的成熟慢病毒颗粒感染CNE1细胞。经流式细胞仪筛选后,建立稳定表达ebv-miR-BART7鼻咽癌细胞株。最后用荧光定量PCR检测其表达水平。结果 PCR和测序验证pLVTHM/BART7重组质粒构建成功;经293FT细胞病毒包装,感染CNE1细胞后,与阴性对照组和未感染组相比,其表达水平明显升高。结论成功构建了pLVTHM/BART7慢病毒重组质粒,建立了稳定表达ebv-miR-BART7的鼻咽癌细胞株CNE1-pLVTHM/BART7,为研究ebv-miR-BART7在鼻咽癌发生发展过程中的基因调控和相应的作用机制提供了有用的细胞模型。
Objective To establish a nasopharyngeal carcinoma（NPC） cell line CNE1-pLVTHM /BART7 with stable ebv-miR-BART7 overexpression.Methods？？The recombinant lentivirus pLVTHM/BART7 expression plasmid was packaged into mature lentivirus by 293FT cells and used to infect CNE1 cells.Flow cytometry was employed for sorting the GFP＋ cells.The efficiency of ebv-miR-BART7 overexpression was determined using qRT-PCR.Results The recombinant lentivirus plasmid pLVTHM/BART7 was successfully constructed and verified by PCR and sequencing.The expression of ebv-miR-BART7 in CNE1 cells infected with the lentivirus pLVTHM/BART7 was significantly increased as compared with the negative control and the blank control cells.Conclusion The recombinant lentivirus vector pLVTHM/BART7 results in high and stable expression of ebv-miR-BART7 in infected CNE1 cells,which provides a useful cell model for further studies of the role of ebv-miR-BART7 in nasopharyngeal carcinoma.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2011年第3期419-422,共4页
Journal of Southern Medical University
基金
国家自然科学基金(3087285630973292)~~
