稳定表达EGFP的不同转移特性骨肉瘤细胞亚株建立及其生物学特性

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pEGFP人人骨肉瘤MG63细胞系，通过细胞电泳，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：发现M6和M8两株均保持人类染色体核型，所形成肿瘤显示人成骨肉瘤上皮样组织形态，其中M6的群体倍增时间为38．4h，软琼脂形成率为18．7％，M8群体倍增时间为23．0h，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。

Isolation and Characterization of Expression of enhanced Green fluorescent Protein Gene in MG63 Cells Subline with Different Metastatic Potential

Object: To study expression of enhanced GFP gene in MG63 cells and isolation and characterization of their subline with different metastatic potential. Methods: after MG63 cells were transfected by enhanced GFP gene in liposome-mediated gene transfer, single cell clones were isolated using limited dilution method. Through preliminary electrophoretic technique and spontaneous metastasis assay in nude mice, two cloned MG63 cell sublines (M6,M8) were selected for further observation on morphology, chromosome number, in vitro growth and invasion assays in nude mice. Results: We found that there was some difference on electrophoretic mobility and in vitro invasive abilities between M6 and M8, and then the population doubling time was 38.4 h and 23.0 h respectively. But there were no obvious differences in other biological feasures among the two sublines. Conclusion: the results show that the MG63 cell line consists of different subpopulations of cells with different metastatic properties. These sublines transfected by EGFP may be valuable for further study on the molecular mechanisms of cancer metastasis.