人成骨细胞株HOS TE85致瘤性转化的研究

目的建立人成骨细胞株HOS TE85致瘤性转化的模型, 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HOS TE85细胞转化浓度后,以MNNG作启动剂,佛波酯(12-0-Te-tradecanoyl phorbol 13-acetate, TPA)作促进剂对其进行转化,66 d后通过细胞形态、软琼脂集落形成实验和裸鼠体内致瘤实验鉴定细胞转化程度.结果经MNNG和TPA协同处理HOS TE85细胞66 d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;转化细胞凝集性增强;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HOS TE85的恶性转化模型.

Neoplastic transformation of human osteoblast-like cell line HOS TE85

Objective To establish human osteoblast-like cell lines TE85 model of neoplastic transformation for exploring molecular mechanism in canceration process of osteosarcoma.Methods HOS TE85 cells were treated by MNNG(initiated factor) and TPA(promotor).The malignancy of transformed cells was identified by observing the cell form,colony forming frequency on soft agar and tumorigenesis in nude mice.Results Continuous passage after induction of neoplastic transformation led to the formation of a few paramorph foci that exhibited an extensively random orientation.The agglomeration in experiment group was more than that in control group.As compared with that of negative control cells,colony formation efficiency of transformed cells in semisolid agar showed a significant increase and the transformed cells could form tumor subcutaneously in the nude mice.The tumors were a poorly differentiated osteosarcoma confirmed by histopathological examination.Conclusion Simulating the process of malignant transformation of human cells,we establish neoplastic transformation of human osteoblast-like cell lines TE85 model.