抑制VEGF-B表达对人绒毛膜癌裸鼠移植瘤影响的研究

目的:研究RNA干扰下调血管内皮生长因子-B(VEGF-B)表达对人绒毛膜癌裸鼠移植瘤生长、侵袭转移及血管生成的影响。方法:将构建的含VEGF-B shRNA的质粒转染人绒毛膜癌细胞JEG-3,建立裸鼠皮下移植瘤模型。观察shRNA干扰VEGF-B表达对JEG-3裸鼠移植瘤生长及侵袭转移能力的影响,并采用免疫组化技术检测移植瘤组织中VEGF-B的表达变化及其所含CD34标记的肿瘤微血管密度(MVD)的差异。结果:(1)抑制VEGF-B表达可减小裸鼠皮下肿瘤大小(P0.05),并降低肿瘤细胞肺转移的发生率(P0.05);(2)免疫组化结果提示,VEGF-B shRNA转染组移植瘤中VEGF-B表达及MVD均低于空白对照组和阴性对照组(P0.05);(3)相关性分析显示,VEGF-B蛋白表达水平与绒毛膜癌MVD高低成正相关。结论:VEGF-B基因在绒毛膜癌的体内生长、侵袭转移中起重要作用,shRNA下调其表达能抑制绒毛膜癌细胞的体内生长、侵袭转移,可能与其调控瘤组织内的微血管形成有关。

The effects of VEGF-B by inhibiting its expression in human chorionic carcinoma transplantation tumor in nude mouse

Objective: To investigate the effect of VEGF-B shRNA on the growth,invasion,metastasis and angiogenesis of the human choriocarcinoma transplanted in nude mice.Methods: We transfected VEGF-B shRNA to human chorionic carcinoma cell line JEG-3,established its subcutaneous transplantation tumor model in a nude mice,and observed the effects of VEGF-B shRNA on growth,invasion and metastasis of JEG-3 cell in vivo,also detected the expression of VEGF-B and CD34 marked tumor microvascular density( microvessel density,MVD) of transplanted tumor tissue by using the technology of immunohistochemical. Result:( 1) As found in JEG-3 subcutaneous transplantation tumor model in nude mice,with inhibiting the expression of VEGF-B,the subcutaneous tumor size in nude mice of VEGF-B shRNA group,as well as the number of pulmonary metastasis cancer nodules,was relatively reduced than the negative control group and the blank group( P < 0. 05).( 2) Immunohistochemical results indicate the expression level of VEGF-B and MVD level were lower in the subcutaneous transplantation tumor of VEGF-B shRNA transfection group than the negative control group and the blank group( P < 0. 05);( 3) Correlation analysis showed that VEGF-B protein expression levels of choriocarcinoma has a positive correlation with its MVD level. Conclusion: VEGF-B gene plays an important role in the growth,invasion and metastasis of choriocarcinoma,inhibition of its expression by shRNA can reduce the growth,invasion and metastasis of JEG-3 cells in vivo,which may be related to its regulation of tumor microvascular formation.