永生化人食管鳞状上皮细胞系的建立及其染色体不稳定性的研究  

Establishment of Immortalized Human Esophageal Squamous Cell Line and Its Chromosomal Instability

作　　者：张浩[1] 蔡昌枰[1] 王士礼[1] 陈学明[1] 叶燕芬[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院,上海200025

出　　处：《肿瘤学杂志》2008年第4期245-250,共6页Journal of Chinese Oncology

摘　　要：[目的]建立永生化人食管鳞状上皮细胞系,并研究细胞永生化过程中以及食管肿瘤细胞染色体的不稳定性。[方法]用人乳头状瘤病毒16型E6/E7(HPV16E6/E7)转染正常人食管鳞状上皮细胞,连续传代培养,检测HPV16E6/E7基因的整合情况及其表达,并对不同代次永生化细胞及食管肿瘤细胞进行端粒荧光原位杂交(FISH)检测和分裂后期细胞形态分析。[结果]HPV16E6/E7基因转染至人食管鳞状上皮细胞后,增殖旺盛,已传至第75代。在细胞危机期(第13和第17代),TTAGGG端粒缺失数为6.0±1.2和4.7±1.5(缺失率分别为6.7%和5.3%),细胞间桥的出现率为28.0%￣30.9%。细胞永生化以后(第47代),端粒缺失数减至2.6±0.7(缺失率3%),细胞间桥出现率降至3.6%,与危机期的比较差异有统计学意义,而与肿瘤细胞的分裂异常相一致。在细胞永生化过程中,细胞间桥的出现率与端粒的缺失率呈正相关性。[结论]HPV16E6/E7基因可以导致人食管鳞状上皮细胞永生化,在此过程中存在较多端粒缺失和细胞间桥,表明染色体的不稳定性在细胞永生化和恶性转变过程中起到重要作用。[Purpose] To establish immortalized esophageal squamous epithelial cell line and to study the chromosomal instability involved in the process of immortalization. [Methods] HPV16 E6/E7 was transfected into esophageal epithelial cells and subcuhured. The expression and integration of E6 and E7 gene was identified. Chromosomal instability was investigated by telomeric FISH. Mitotic cell morphology was analysed during the cell immortalization. [Results] HPV16 E6/E7 transfected esophageal cell line was subcuhured to the 75th generation and proliferation was exuberant. During cell crisis stage （13rd and 17th generation）, TYAGGG telomere deletion was 6.0±1.2 and 4.7±1.5 （deletion rate about 6.7% and 5.3%）. The occurrence of anaphase bridges were 28.0%-30.9%. After immortalization （47th generation）, telomere deletion reduced to 2.6±0.7 （deletion rate about 3%）, the occurrence of anaphase bridges was 3.6%, which was significant difference with crisis stage, in accordance with mitotic abnormality occurred in esophageal neoplasma cell. The frequency of anaphase bridge was positively correlated with that of telomere deletion during immortalization. [Conclusion] HPV16 E6/E7 may result in immortalization of esophageal epithelial cells, and cause telomere dysfunction and consequent anaphase bridges existed in the process of immortalization. Chromosomal instability may play an important role of cell immortalization and carcinogenesis.

关 键 词：上皮细胞 染色体 肿瘤 实验性 食管肿瘤 

分 类 号：R73-35[医药卫生—肿瘤]