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目的建立经绿色荧光蛋白基因转染的骨肉瘤细胞亚株并研究其生物特性。方法利用脂质体转染增强型绿色荧光蛋白真核表达质粒(pEGFP-N1)人人骨肉瘤MG63细胞系,通过有限稀释法和细胞电泳,获得两株细胞克隆M6和M8,经体外细胞增殖、软琼脂形成、生长曲线、裸鼠成瘤试验综合分析其生物学行为改变。应用组织形态学观察、染色体分析、软琼脂克隆形成法研究癌细胞的生物学特性。结果M6和M8两株在细胞电泳率和侵袭性上差异有统计学意义(P＜0．05),其中M6的群体倍增时间为38．4 h,软琼脂形成率为18．7%,M8群体倍增时间为23．0 h,软琼脂形成率为29．3%。裸小鼠背部皮下接种M6和M8,发现M8成瘤时间短,细胞增殖快,但在4周内两者均不发生转移。结论骨肉瘤细胞亚系有不同的转移特性,GFP的整合及表达未对MG63细胞的生长状态造成明显影响,可作为报告基因进一步了解骨肉瘤细胞转移的差异性分析。
Objective To establish a sub-strain of osteosarcoma cells transfected with green fluorescent protein gene and study its biological characteristics. Methods The human eukaryotic expression plasmid pEGFP-N1 (pEGFP-N1) was transfected into human osteosarcoma MG63 cell line by Lipofectamine 2000. Two cell clones M6 and M8 were obtained by limiting dilution and cell electrophoresis. Soft agar formation, growth curve, tumor formation test in nude mice comprehensive analysis of its biological behavior changes. Morphological observation, chromosome analysis, and soft agar cloning method were used to study the biological characteristics of cancer cells. Results The electrophoretic rate and invasiveness of M6 and M8 strains were significantly different (P<0.05). The population doubling time of M6 was 38.4 h, and the soft agar formation rate was 18.7%. M8 The population doubling time was 23.0 h and the soft agar formation rate was 29.3%. M6 and M8 were inoculated subcutaneously on the back of nude mice. M8 tumor formation time was short and cell proliferation was rapid, but no metastasis occurred within 4 weeks. Conclusions Osteosarcoma cell sublines have different metastasis characteristics. The integration and expression of GFP have no significant effect on the growth of MG63 cells. It can be used as a reporter gene to further understand the differential analysis of osteosarcoma cell metastasis.