人永生化成骨细胞hFOB1.19恶性转化的研究  

Malignant transformation of immortalized human fetal osteoblastic cell line hFOB1.19

作　　者：孟刚[1] 李懿[1] 辛榕[1] 郭乔楠[1] 

机构地区：[1]第三军医大学西南医院病理学研究所,重庆400038

出　　处：《第三军医大学学报》2007年第5期380-383,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30371446)~~

摘　　要：目的建立人胚永生化成骨细胞系(hFOB1.19)的恶性转化模型,作为骨肉瘤癌变过程细胞分子生物学研究的模型。方法通过克隆形成率实验确定N-甲基-N’-硝基-N-亚硝基胍(N-methyl-N’-nitro-N-nitrosoguanidine,MNNG)对hFOB1.19细胞转化浓度后,以MNNG作启动剂,佛波酯(12-0-Te-tradecanoyl phorbol 13-acetate,TPA)作促进剂对hFOB1.19细胞进行转化,90d后通过细胞形态、DNA含量分析、软琼脂集落形成试验和裸鼠体内致瘤试验鉴定细胞恶性转化程度。结果经MNNG和TPA协同处理hFOB1.19细胞90d后,细胞中出现形态异常的转化灶,转化灶细胞失去接触抑制;高倍体细胞数,转化组(81.08%)高于对照组(55.03%);软琼脂克隆形成率明显增加;转化细胞在裸鼠皮下成瘤,病理组织学证实为低分化骨肉瘤。结论模拟人体细胞发生恶性转化的过程,建立了人胚永生化成骨细胞系的恶性转化模型。Objective To establish a malignant transformed human fetal osteoblastic human cells from the immortalized cell line hFOB1.19 in order to explore the molecular mechanism in tumorigenesis of osteosarcoma. Methods hFOB1.19 cells were treated sequentially by an initiated factor, N-methyl-N＇-nitro-N-nitrosoguani- dine （MNNG） and a promoter, 12-O-tetradecanoyl phorbol-13-acetate （TPA）. The features of malignancy of transformed cells were identified by cell morphology, DNA content analysis, colony forming frequency on soft agar and tumorigenetic test in nude mice. Results Continuous passaging after the treatments resulted in the formation of a few paramorph foci and exhibited in an extensively random orientation. The poly-ploid of DNA was 81.08% in experiment group, much higher than that in control group （55.03%）. Compared with that of negative control cell, colony formation efficiency of transformed cells in semisolid agar showed a a significant increase. The transformed cells formed tumors subcutaneously in the nude mice, which were verified to be poorly differentiated osteosarcoma histopathological examination. Conclusion Mocking the process of malignant transformation of human cells, we establish malignant transformed immortalized human fetal osteoblastic human cells （hFOB1.19） model.

关 键 词：人永生化成骨细胞 MNNG TPA 恶性转化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R73-354[医药卫生—基础医学]