人类风湿性关节炎滑膜细胞系(RASB)的建立和生物学特性的观察  

ESTABLISHMENT AND CHARACTERIZATION OF A HPV16E6/E7 TRANSFORMED HUMAN RHEUMATOID ARTHRITIS SYNOVIAL CELL LINE RASB

作　　者：蒋知新[1] 蒋明[2] 陈志强[2] 张焕相[1] 彭安[1] 王永潮 

机构地区：[1]北京师范大学细胞增殖及调控生物学教育部重点实验室 [2]中国医学科学院北京协和医院,北京100005

出　　处：《解剖学报》2002年第3期268-273,共6页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目 ( 39430 0 80 ) ;国家重点基础研究项目 (G1990 5 390 1)

摘　　要：目的　建立人类风湿性关节炎滑膜细胞永生细胞系。　方法　用重组有HPV16病毒E6 E7基因阅读框架的逆转录病毒载体转染原代培养的人类风湿性关节炎滑膜细胞 ,经G418筛选 ,获取细胞克隆RASB ,并从形态学、生长特性、核型组成、致瘤性和分泌功能等多方面对建系细胞RASB进行生物学观察。　结果　实验和观察证实 ,转化滑膜细胞染色体整合HPV病毒DNA ,表达HPVE6蛋白 ,基本保留了B型滑膜细胞特征形态、细胞骨架和分泌功能 ,倍增时间缩短一半 ,对裸鼠无致瘤性 ,软琼脂培养形成细小集落。已稳定传代大于 10 0代。　结论　建立了能长期体外稳定传代的人类风湿性关节炎滑膜细胞系。此细胞系的建立将为类风湿关节炎致病机理的研究和治疗提供极有意义的体外细胞模型。Objective To establish human rheumatoid arthritis synovial cell line. Methods Primary rheumatoid synovial cells were transfected with recombinant retroviral vector containing the E6 and E7 open reading frames of HPV 16.Individual cell clone RASB was selected in the medium supplemented with G418.Both transformed rheumatoid synovial cells and nontransformed cells were observed by means of morphology,growth property,chromosomal analysis,tumorigenicity,and secretive function. Results The transformed synovial cells expressed HPV16 E6/E7 genes and E6 protein by PCR and Western blotting.The transformed cells maintained similar morphology,cytockeleton and secretive function of type B synovial cell.The doubling time of RASB cells were decreased to one half of non-transformed ones.The cell line was not tumorigenic in nude mice but formed small cell colonies in soft agarose.These cells have being in culture more than 100 passages.In contrast,the primary rheumatoid synovial cells did not survive the culture environment beyond 20 to 30 passages.Conclusion We have established a permanent type B rheumatoid synovial cell line successfully.The established cell line might be served as a very useful model in studying of rheumatoid arthritis.;

关 键 词：类风湿性关节炎 滑膜细胞 人乳头瘤病毒16型 基因表达 细胞系 

分 类 号：R593.22[医药卫生—内科学]