文档介绍：PAR-2基因在EBV转化人胃上皮细胞中的作用
刘换新陈娟刘梅叶春
【摘要】目的研究EBV感染人胃上皮细胞系GFS-1后PAR-2基因的表达状况及转化作用,以探讨PAR-2基因在Epstein-Baar病毒(EBV)相关胃癌发生中所起的作用。方法用携带NEO基因的重组EBV产生细胞系Akata 1061以密切接触法感染人胃上皮细胞系GF5-1,采用脂质体介导法将PAR-2基因转染至同一细胞,以pcDNA3空载体质粒转染同一细胞为对照;经G418筛选获得感染或转染的抗性细胞克隆;采用免疫细胞化学法测定EBNA1的表达以鉴定EBV感染细胞克隆,测定EBV感染和PAR-2基因转染细胞中PAR-2蛋白的表达;通过形态学观察、细胞生长曲线测定及流式细胞分析等方法观察细胞生物学特性的变化。结果与对照组相比,EBV感染及PAR-2转染后细胞发生明显的形态学变化,生长速度明显加快,S期细胞比例显著提高,分别为(±)%、(±) % 和(±)%,差异有显著性(P<),表明EBV感染及PAR-2基因转染均使细胞增殖加快。结论EBV感染及PAR-2基因转染可使人胃上皮细胞的增殖速度加快,细胞恶性程度增强,但并未导致肿瘤的发生。EBV对人胃上皮细胞的转化作用可能与PAR-2基因的过度表达有关。
【关键词】 Epstein-Barr 病毒(EBV);PAR-2;人胃上皮细胞;基因转染;细胞周期
Role of PAR-2 gene in EBV-transformed human gastric epititelial cells
LIU Huan-xin,CHEN Juan,LIU Mei,et Hospital of Armed Police of Guangdong,Guangzhou 510507,China
[Abstract]ObjectiveTo define the role of PAR-2 gene in the carcinogenesis of EBV-associated gastric carcinoma, we examined the expression of PAR-2 in the human epithelial cell line GES1 infected with EBV and tested if EBV could induce cell 1061 was used to infect GES-1 the intimate contact meth-od. pcDNA3-PAR-2 was hansfected into cells the UpofectAMINE method, whereas pcDNA3 vector was used as the control. icin 418 (G418) was adopted to select positive clones that were either EBV or PAR-2 positive. Immuno-histochemical staining method was used to detect EBNAI expression in cell clones infected with EBV and the expressionof PAR-2 protein in EBV-infected or PAR-2-transfected cell clones. Morphological observations, cell growth curve de-terminations, soft agar gel formation tests, and flow cytometry were employed to measure changes in cell proliferationand cell cycle to the control, EBV-infected or PAR-2-transfected cells revealed significant morphological changes that were panied a significant increase in the cell growth velocity. The numbers of S ase cells were significantly increased(P<), both EBV-infection (
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