自发永生性转化人胚肺上皮细胞系HLEC的建立及鉴定  

ESTABLISHMENT AND CHARACTERIZATION OF A SPONTANEOUSLY IMMORTALIZED HUMAN LUNG EPITHELIAL CELL LINE HLEC

作　　者：张志谦[1] 胡颖[1] 王冰晶[1] 赵威[1] 林仲翔[1] 

机构地区：[1]北京大学临床肿瘤学院暨北京市肿瘤防治研究所细胞生物室,北京100034

出　　处：《解剖学报》2005年第2期113-116,共4页Acta Anatomica Sinica

基　　金：国家重大基础研究发展规划项目"973"课题(G2000057002);国家自然科学基金资助项目 (30270658;30370555);北京市自然科学基金重点项目(7021001);北京大学"985"项目;北京大学"十五""211"工程重点学科建设项目资助(534)

摘　　要：目的　对源自人胚肺组织原代培养、形态似上皮类型的细胞系———人肺上皮细胞(HLEC)的生物学特 性进行鉴定,为相关研究和应用提供模型和依据。　方法　分别采用相差显微镜下观察、细胞计数、β 半乳糖苷酶 染色、免疫荧光细胞化学。人Alu序列PCR扩增、染色体计数、软琼脂集落形成实验和裸鼠致瘤实验对HLEC的形 态学特征、生长特性、衰老状态、上皮细胞标志物、来源、细胞遗传学特点和恶性转化等生物学特性进行研究鉴定。 　结果　HLEC角蛋白和上皮型细胞间黏附分子钙黏着素E cadherin免疫荧光染色阳性,染色体众数在41～44,人 Alu序列PCR扩增结果阳性,β 半乳糖苷酶阳性细胞的比例未见随细胞代数的增加而升高,软琼脂和裸鼠体内均不 生长,微丝及黏着斑结构正常。　结论　HLEC细胞具有永生化细胞特征、属相对正常人肺上皮亚二倍体细胞,可 用于肺癌癌变机理研究和癌细胞的相对正常对照。Objective The aim of this study was to characterize the biological natures of human lung epithelial cells(HLEC), which were subcultured from human fetal lung primary culture. Methods HLEC cells were assayed for their growth characters by measuring population doubling time and soft-agar colony forming experiment. Epithelial molecular markers were detected by keratin and E-cadherin immunofluorescent staining. Chromosome medal number and human Alu-sequence based PCR were also performed to characterize their cytogenetic nature and specie origin. The cells were further inoculated into nude mice to exclude tumorigenecity. Finally, β-galactosidase staining was used for evaluating its senescence status. Results HLEC cells had constant population doubling time among different passages, and very few β-galactosidase positive cells were found even in cultures over 100 passages. They were positive for keratin, E-cadherin, with normal structure of microfilaments. Chromosome(medal number was 41-44) and Alu-PCR verified its human origin. Furthermore, they were not tumorigenetic in nude mice. Conclusion HLEC cells are spontaneously immortalized, sub-diploid human lung epithelial cells. They can be used as a model for the research of lung cancer.

关 键 词：人肺上皮细胞 不死性转化 钙黏着素 角蛋白 亚二倍体 

分 类 号：Q249[生物学—细胞生物学] R37-73[医药卫生—病原生物学]