c-myc基因在EBV转化人胃上皮细胞中的作用  

Role of c-myc geae in EBV-induced transformaion of human gastric epithelial cells

作　　者：习瑾昆[1] 周天戟[3] 杨方[1] 李治国[1] 朱丽华[2] 

机构地区：[1]华北煤炭医学院实验中心,唐山063000 [2]华北煤炭医学院生物技术系,唐山063000 [3]江苏大学医学技术学院微生物学教研室,212013

出　　处：《中华微生物学和免疫学杂志》2007年第3期202-206,共5页Chinese Journal of Microbiology and Immunology

基　　金：教育部留学回国人员科研启动基金（2000479）;志谢：十分感谢日本北海道大学医学部遗传医学研究所肿瘤病毒学部Takada教授提供Akata 1061细胞系、pcDNA3和pcDNA3-c-myc质粒,感谢华北煤炭医学院附属医院耳鼻喉科提供鼻咽癌病人血清.

摘　　要：目的研究EBV感染人胃上皮细胞系GES-1后c-myc基因的表达状况及转化作用，以探讨c-myc基因在Epstein-Barr病毒（EBV）相关胃癌发生中所起的作用。方法用携带NEO^r基因的重组EBV产生细胞系Akata1061以密切接触法感染人胃上皮细胞系GES-1，采用脂质体介导法将c-myc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EBNAl的表达以鉴定EBV感染细胞克隆，测定EBV感染和c-myc基因转染细胞中C．myc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c-myc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（70．96±0．91）％和（60．67±3．06）％vs（34．74±1．03）％，P〈0．05，表明EBV感染及c-myc基因转染均使细胞增殖加快。结论EBV感染及c-myc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c-myc基因的过度表达有关。Objective To define the role of c-myc gene in the carcinogenesis of EBV-associated gastric carcinoma, we examined the expression of c-myc in the human epithelial cell line GES-1 infected with EBV and tested if EBV could induce cell transformation. Methods Akata 1061, was used to infect GES-1 by the intimate contact method. pcDNA3-c-myc was transfected into cells by the LipofectAMINE method, whereas pcDNA3 vector was used as the control. Geneticin 418 （G418） was adopted to select positive clones that were either EBV or C-myc positive. Immunohistochemical staining method was used to detect EBNA1 expression in cell clones infected with EBV and the expression of C-myc protein in EBV-infected or c-myc-transfected cell clones. Morphological observations, cell growth curve detenninafions, soft agar gel formation tests, and flow cytometry were employed to measure changes in cell proliferation and cell cycle progress. Results Compared to the control, EBV-infected or c-myc-transfected cells revealed significant morphological changes that were accompanied by a significant increase in the cell growth velocity. The numbers of S-phase cells were significantly increased （P〈0.05） by both EBV-infection （70.96% ±0.91%） and c-myc-transfection （60.67% ± 3.06% ）, compared to the control （34.74% ± 1.03% ）. Cell colony formation was not detected by the soft agar gel clone formation test. Conclusion Both EBV infection and c-myc transfection promote the proliferation of gastric epithelial cells result in an increased tendency of malignance. However, the both events failed to induce carcinoma. These data suggest that an over-expression of c-myc may play a role in EBV induced transformation on human gastric epithelial cells.

关 键 词：Epstein-Barr病毒(EBV) C-MYC 人胃上皮细胞 基因转染 细胞周期 

分 类 号：R686[医药卫生—骨科学]