EB病毒介导人鼻咽上皮细胞永生化早期阶段的生物学观察

OBSERVATION OF THE BIOLOGICAL CHARACRERIZATIONS OF NASOPHARYNGEAL EPITHELIAL CELLS BY EB VIRUS INFECTION IN EARLY PHASE OF IMMORTALIZATION

EB病毒转化人鼻咽上皮细胞,建立体外多阶段细胞模型有利于从细胞和分子水平对肿瘤发病机制作深入研究。我们利用与鼻咽癌密切相关的EB病毒和TPA的协同作用,观察原代人胚鼻咽上皮细胞逃避老化期后其生物学特性的变化。结果表明,EB病毒感染的人胚鼻咽上皮细胞在原代培养后期老化相关半乳糖苷酶(SA-β-Gal)表达降低,形态学上发生改变,出现转化灶样集落,群体倍增时间降低,体外培养寿命延长,表明EB病毒促使部分原代人胚鼻咽上皮细胞逃避老化期、进入永生化早期阶段。这些研究资料为进一步阐明上皮细胞永生化分子机制及建立人鼻咽上皮细胞永生化模型提供实验依据。

The multi-stage cell model of the nasopharyngeal carcinoma development in vitro by Epstein-Barr virus transformation is beneficial for the elucidation of the mechanism of nasopharyngeal cancer. To observe the biological changes of primary human nasopharyngeal epithelial cells in early phase of immortalization, in this study, we have detected the morphological changes and the expression profile of senescence-associated β-galactosidase (SA-β-Gal) in primary culture. In addition, the expression of EB virus latent membrane protein 1 (LMP1) and the growth curve of primary cells were also detected. Our results showed a low percentage of cells infected with EB virus expressing SA-β-Gal activity at the late primary culture. In morphology , the cells also formed multilayer foci, and the cell population doubling time was showed. These results demonstrated that the nasopharyngeal epithelial cells by EB virus infection have passed through the senescence and entered the early phase of immortalization. These cells have some of the transformed characteristics. Our results provided the data for further study on the mechanism of immortalization and the establishment of human nasopharyngeal epithelial cell line.