Epstein-Barr virus induces human nasopharyngeal epithelial cells to escape from the replicative senescence  

EB病毒诱导人鼻咽上皮细胞逃避老化期的生物学研究(英文)

作　　者：杨静[1] 唐发清[1] 顾焕华[1] 邓锡云[1] 翁新宪[1] 唐敏[1] 曹亚[1] 

机构地区：[1]湖南医科大学肿瘤研究所,长沙410078

出　　处：《Chinese Medical Journal》2002年第6期803-809,146,共7页中华医学杂志（英文版）

基　　金：grantsfromStateKeyProgramofBasicResearch (G19980 5 12 0 1)andNationalScienceFundforDistinguishedYoungScholars (No . 395 2 5 0 0 2 2 )fromNationalNaturalScienceFoundationofChina ;andCMB (No .96 6 5 5 )fromtheChinaMedicalBoardofNewYork ;USA

摘　　要：OBJECTIVE: To observe the biological changes of primary human nasopharyngeal epithelial cells in the early stage of immortalization. METHODS: The morphological changes of nasopharyngeal epithelial cells were observed by phase contrast microscopy, and the activity profile of senescence-associated beta-galactosidase (SA-beta-Gal) was detected by SA-beta-Gal staining. The expression of p16(INK4a) protein was tested by immunochemical assay, and the life span in vitro of nasopharyngeal epithelial cells was calculated as population doublings. In addition, the expression of Epstein-Barr (EB) virus latent membrane protein 1 (LMP1) was also detected by immunofluorescence staining. RESULTS: Morphologically, cells treated with EB virus and 12-o-tetradecanoyl-phorbol-13-acetate (TPA) formed multi-layer foci, and their cellular life span in vitro was extended (about 155 days of culture). A low percentage of cells (about 4.8%) expressed SA-beta-Gal activity at late primary culture, and did not always express p16(INK4a) protein in the progression of culture. CONCLUSIONS: Nasopharyngeal epithelial cells treated with EB virus in cooperation with TPA can pass through the stage of senescence and enter the early stage of immortalization. Some changes of phenotype occur in these cells.Our results provide data for further studying the mechanism of immortalization and the establishment of a human nasopharyngeal epithelial cell line.Objective To observe the biological changes of primary human nasopharyngeal epithelial cells in the early stage of immortalization. Methods The morphological changes of nasopharyngeal epithelial cells were observed by phase-contrast microscopy, and the activity profile of senescence-associated β-galactosidase (SA-β-Gal) was detected by SA-β-Gal staining. The expression of p16 INK4a protein was tested by immunochemical assay, and the life span in vitro of nasopharyngeal epithelial cells was calculated as population doublings. In addition, the expression of Epstein-Barr (EB) virus latent membrane protein 1 (LMP1) was also detected by immunofluorescence staining. Results Morphologically, cells treated with EB virus and 12-o-tetradecanoyl-phorbol-13-acetate (TPA) formed multi-layer foci, and their cellular life span in vitro was extended (about 155 days of culture). A low percentage of cells (about 4.8%) expressed SA-β-Gal activity at late primary culture, and did not always express p16 INK4a protein in the progression of culture. Conclusions Nasopharyngeal epithelial cells treated with EB virus in cooperation with TPA can pass through the stage of senescence and enter the early stage of immortalization. Some changes of phenotype occur in these cells. Our results provide data for further studying the mechanism of immortalization and the establishment of a human nasopharyngeal epithelial cell line.

关 键 词：Cell Aging Cell Transformation  Viral Cyclin-Dependent Kinase Inhibitor p16 Epithelial Cells Herpesvirus 4  Human Humans NASOPHARYNX Research Support  Non-U.S. Gov't Tetradecanoylphorbol Acetate 

分 类 号：R766.3[医药卫生—耳鼻咽喉科]