人LYVE-1基因联合小鼠GMC-SF基因干预小鼠膀胱癌的生长和转移

Antitumor effects of interference of xenogeneic human LYVE-1 gene combined with mouse GMC-SF gene on growth and metastasis of bladder cancer in mice

目的:研究人淋巴管内皮细胞透明质酸受体-1(lymphatic vessel endothelial hyaluronan receptor-1,LYVE-1)联合小鼠粒细胞巨噬细胞集落刺激因子(granulocyte macrophage colony-stimulating factor,GMC-SF)干预小鼠膀胱癌的生长和转移。方法:通过人和小鼠的肺组织扩增LYVE-1基因的cDNA,构建真核表达载体phLYVE-1和pmLYVE-1,采用脂质体转染体外培养的小鼠膀胱移行细胞癌BTT-T739细胞、小鼠成纤维细胞株NIH-3T3和人胚肾细胞株AD293,用电穿孔法转染小鼠肌肉组织。采用Western印迹法验证LYVE-1蛋白的表达。用电穿孔法将phLYVE-1联合pmGMC-SF导入T739小鼠肌肉内,1周后皮下接种膀胱移行细胞癌BTT-T739-gfp细胞,观察肿瘤生长、淋巴结转移及全身转移情况。结果:成功构建真核表达载体phLYVE-1和pmLYVE-1。经Western印迹法检测证实,phLYVE-1和pmLYVE-1能在被转染的细胞和小鼠肌肉组织中有效表达。预先接受了phLYVE-1和pmGMC-SF的小鼠,BTT-T739移植瘤的生长减慢,肿瘤体积小于对照组,且肿瘤肺转移明显减少,肿瘤附近和远处淋巴结增大并呈现明显的增生反应。结论:异种同源LYVE-1基因联合小鼠GMC-SF基因干预对小鼠膀胱移行细胞癌的生长和转移有抑制作用。

Objective： To study the effect of interference of xenogeneic human lymphatic vessel endothelial hyaluronan receptor-1（LYVE-1） gene combined with granulocyte macrophage colony-stimulating factor（GMC-SF） gene on growth and metastasis of bladder cancer in mice.Methods： The cDNA of LYVE-1 gene was amplified from human and mouse lung tissues.Mammalian eukaryotic expression vectors phLYVE-1 and pmLYVE-1 were constructed.The vectors were transfected into mouse transitional bladder cancer cells BTT-T739,mouse fribroblast NIH-3T3 cells,and HEK-AD293 cells.The vectors were transfected in mouse muscle tissues in vivo by electroporation method.LYVE-1 protein expression was verified by Western blotting.The vectors of phLYVE-1 combined with mouse pmGMC-SF were electroporated into muscle tissues of the legs of mice,then bladder cancer cells BTT-T739-gfp were subcutaneously implanted into mice one week later.The tumor growth and lymph node and whole body metastasis were observed.Results： The phLYVE-1 and pmLYVE-1 vectors were successfully constructed.PhLYVE-1 and pmLYVE-1 were efficiently expressed in transfected cells and mouse muscle tissues.The mice that received phLYVE-1 and pmGMC-SF exhibited delayed growth of xenografted tumor and relatively small tumor size.The pulmonary metastasis significantly decreased,and the adjacent and distant lymph node showed apparent proliferation reaction.Conclusion：Simultaneous interference of xenogeneic LYVE-1 gene and mouse GMC-SF gene inhibit the growth and metastasis of transitional bladder cancer in mice.