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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 investigate the role of EBV-encoding BARF1 gene in the malignant transformation of human gastric epithelial cells. METHODS: Human gastric epithelial cell line GES-1 was transfected with the eukaryotic vector PIRES2-EGFP / BARF1 carrying BARF1 gene and screened by G418 to obtain human gastric epithelial cell line stably expressing BARF1 gene. GES-1 cells were divided into four groups, including one cell line transfected with BARF1 gene, one transfected with BARF1 gene and TPA-stimulated cell group, one transfected empty vector cell group and untransfected cell group. The colony formation ability of soft agar and the ability of tumorigenesis in SCID mice were observed. SCID mice were divided into different groups for subcutaneous inoculation, 3 in each group. Results The colonies transfected with BARF1 gene and their TPA - stimulated groups could form clones in soft agar. The colony formation rate was 24.2% (58/240) in the transfected group and 40.0% (96/240) in the TPA group. However, none of the untransfected cells and empty vector transfected cells formed no colonies in soft agar. The cells transfected with BARF1 gene could form tumors in SCID mice. The untransfected cells transfected with empty vector and transfected with BARF1 gene failed to form tumors in SCID mice, but the transfected BARF1 gene Of the cell groups produced nodules in SCID mice. Conclusions BARF1, an oncogene of EBV, can cause malignant transformation of cells and acquire the characteristics of tumor cell growth. It plays an important role in the malignant transformation of human gastric epithelial cells.