摘要
目的探讨EB病毒编码BARF1基因在人胃上皮细胞恶性转化中的作用。方法用携带BARF1基因的真核载体PIRES2-EGFP/BARF1,转染人胃上皮细胞系GES-1,通过G418筛选,获得稳定表达BARF1基因的人胃上皮细胞株。GES-1细胞被分为4组,包括1个转染BARF1基因的细胞组,1个转染BARF1基因并有TPA刺激细胞组,1个转染空载体细胞组和未转染的细胞组。观察软琼脂克隆形成能力和SCID鼠体内成瘤能力。SCID鼠被分成不同的组进行皮下接种,每一组接种3只。结果转染BARF1基因的细胞组及其TPA刺激组在软琼脂中能形成克隆,转染组克隆形成率为24.2%(58/240),TPA组克隆形成率为40.0%(96/240);而未转染组和转染空载体组细胞在软琼脂中没有克隆形成。转染BARF1基因的TPA刺激组细胞在SCID鼠体内能形成了肿瘤;未转染组、转染空载体组及转染BARF1基因的细胞组在SCID鼠体内没能形成肿瘤,但转染BARF1基因的细胞组在SCID鼠体内产生了结节。结论作为EBV的一个癌基因BARF1能够使细胞发生恶性转化,获得肿瘤细胞生长的特性,在人胃上皮细胞恶性转化过程中起重要作用。
Objective To explore the role of BARF1 gene encoded by Epstein-Barr virus（EBV） in malignant transformation of human gastric epithelial cells.Methods BARF1 gene carried by the eukaryotic vector pIRES2-EGFP/BARF1 was transfected into human gastric epithelial cell line GES-1;the transfected cells were screened by G418;and through this process obtained human gastric epithelial cells that stably expressed BARF1 gene.The colony formation in soft agar and tumorigenicity in SCID mice of the stably transfected cells were examined.GES-1 cells were divided into 4 groups including a BARF1 group（cells transfected with BARF1 gene）,a TPA group（cells transfected with BARF1 gene and treated with TPA）,a vector group（cells transfected with vector plasmid） and a control group（cells with no transfection）.The SCID mice were given different groups of cells through subcutaneous inoculation,and each treatment included 3 mice.Results The cells of the BARF1 group and the TPA group formed colonies in soft agar,and the clony-forming efficiency were 24.2%（58/240） and 40.0%（96/240）,respectively,while cells of the vector group and the control group did not form any colonies in soft agar.Tumors were formed in the SCID mice that were given the cells of the TPA group,while no tumor was formed in the SCID mice that were given the cells of the other groups.However,nodules were formed in the SCID mice that were given the cells of the BARF1 group.Conclusion BARF1 gene,as an oncogene of EBV,can induce malignant transformation and gain of tumor cell characteristics in cells and plays an important role in the malignant transformation of gastric epithelial cells.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2011年第19期2025-2028,共4页
Journal of Third Military Medical University
基金
河北联合大学博士启动基金
传染病预防控制国家重点实验室自主研究重点课题(2011SKLID103)~~
关键词
人胃上皮细胞
转染
BARF1基因
human gastric epithelial cells
transfection
BARF1 gene
