摘要
目的:建立表达Epstein-Barr(EB)病毒核抗原-1(EBNA1)的肿瘤细胞株,用于EB病毒相关肿瘤的研究。方法:将1 200 bp的EBNA1基因片段插入真核细胞表达载体pcDNA3中,使用脂质体介导pcDNA3/EBNA1质粒进入人鼻咽癌CNE2细胞和人宫颈癌Hela细胞,通过G418进行阳性克隆的筛选,PCR、RT-PCR进行鉴定。结果:成功建立EBNA1基因阳性的鼻咽癌和宫颈癌肿瘤细胞株,经PCR、RT-PCR检测证明两种转基因细胞株稳定表达EBNA1。结论:EBNA1阳性肿瘤细胞株的建立,为体外进行EB病毒相关肿瘤的研究提供细胞模型。
Purpose: To establish tumor cell strain expressing Epstein-Barr virus nuclear antigen 1 ( EBNA1). Methods: Plasmid pcDNA3/EBNAl was derived from pcDNA3 by inserting the gene which encodes EBNAl fragment and was transfected into human cervical cancer Hela cell and human nasopharyneal cancer CNE2 cell by Lipofectamine Reagent. Clones of cells were screened by G418 and identified by PCR, RT-PCR and Western blot assays. Results: The two transfected EBNA1 gene cell lines consistently expressed the EBNA1 over a long period. Conclusions: The EBNA1 positive tumor cell strains may serve as good tools for research in EBV associated tumors.
出处
《中国癌症杂志》
CAS
CSCD
2004年第2期105-108,共4页
China Oncology
基金
国家自然科学基金重大国际(地区)合作研究(No.30120160823)
国家基金资助项目(No.C030310)
广东省自然科学基金重点项目(No.020100)
广东省自然科学基金(No.010637)
全军十五科研基金(No.01MA135)。
