内皮抑素基因对裸鼠移植骨肉瘤的抑制作用

Inhibitory effect of endostatin gene on transplanted osteosarcoma in nude mice

目的:探讨内皮抑素(endostatin,ES)基因对裸鼠骨肉瘤细胞UMR106移植瘤的抑制作用。方法:携带ES基因重组质粒体外扩增后,应用脂质体法转染骨肉瘤细胞UMR106,Zeocin抗性筛选得到ES-UMR106细胞。MTT法观察ES基因对肿瘤细胞增殖的影响,ELISA法检测转染后肿瘤细胞的ES分泌情况。MTT法观察ES基因对血管内皮细胞增殖能力的影响。应用ES-UMR106和UMR106细胞制作裸鼠皮下移植瘤模型,观察两组动物移植瘤生长情况、瘤组织的病理变化及肺转移的情况。结果:ES基因转染不影响UMR106细胞的增殖能力;ES-UMR106细胞可表达有生物活性的ES,转染后48hES在培养液中超过350ng/ml。ES-UMR106细胞分泌的ES可明显抑制血管内皮细胞增殖。与UMR106细胞相比,ES-UMR106细胞在裸鼠接种的移植瘤生长速度缓慢、包膜完整,病理组织学显示瘤体内血管稀少,肿瘤细胞广泛坏死;UMR106细胞移植瘤裸鼠肺组织出现转移灶(2/8),ES-UMR106细胞移植瘤裸鼠无肺转移灶。结论:携带ES基因重组质粒转染骨肉瘤UMR106细胞后可抑制该骨肉瘤细胞移植瘤的生长和转移。

Objective：To investigate the inhibitory effects of endostatin （ES） on the growth and metastasis of transplanted osteosarcoma UMR106 cells in nude mice. Methods： pBudCE4.1/ES was amplified and transfected into osteosarcoma cell line UMR106 through lipidosome; the non-transfected UMR106 cells were eliminated by Zeocin. The proliferation of ES-UMR106 cells was examined by MTT assay and the production of ES by ES-UMR106 cells was investigated by ELISA. The effects of ES on the in vitro proliferation of vascular endothelial cells were observed by MTT assay. Sixteen nude mice were randomly divided into 2 groups： one group was transplanted with UMR106 cells and the other with ES-UMR106 cells. The tumor size, pathological observation, tumor angiogenesis, and the pulmonary metastasis were observed. Results： The proliferation of UMR106 cells was not affected by ES transfection. ES-UMR106 cells expressed bioactive ES, and the ES level in the medium was higher than 350 ng/ml 48 h after transfection. ES produced by ES-UMR106 cells significantly inhibited the proliferation of vascular endothelial cells. Compared with UMR106 transplanted tumor, ES-UMR106 transplanted tumor grew slowly in nude mice, and the formed tumors was well-margined and had less angiogenesis and mass necrosis. Two of the 8 mice transplanted with UMP106 cells had pulmonary metastasis and no metastasis was found in ES-UMR106 transplanted group. Conclusion： Transfection with recombinant endostatin plasmid can inhibit the growth and metastasis of transplanted osteosarcoma in nude mice by inhibiting vascular endothelial cell proliferation.