反义VEGF_(165)基因转染抑制裸鼠体内人神经母细胞瘤的生长  

Inhibition of growth of human neuroblastoma in nude mouse models by antisense VEGF_(165) cDNA transfection

作　　者：温铭杰[1] 李慎涛[1] 王雅梅[2] 祁雅慧[1] 

机构地区：[1]首都医科大学免疫学系,北京100069 [2]首都医科大学生化与分子生物学系,北京100069

出　　处：《基础医学与临床》2007年第11期1214-1218,共5页Basic and Clinical Medicine

基　　金：北京市教委基金(099KJ-08)

摘　　要：目的研究反义VEGF165cDNA转染对裸鼠移植性人神经母细胞瘤血管生成及超微结构的影响。方法将稳定转染正义VEGF165cDNA、反义VEGF165cDNA及空载体的人神经母细胞瘤细胞株SH-SY5Y接种于裸鼠皮下,观察肿瘤生长情况;HE染色观察肿瘤组织病理变化;透射电镜观察肿瘤组织超微结构;免疫组化染色比较裸鼠移植瘤微血管密度(MVD)。结果与空载体组相比,反义组裸鼠肿瘤瘤块体积小(P<0.05),MVD显著减少(P<0.01),肿瘤组织内血管稀疏,核分裂相少见,内皮细胞变性、肿胀,血液淤滞,凋亡细胞多见。结论反义VEGF165cDNA转染能抑制裸鼠人神经母细胞瘤的生长,降低瘤块内微血管生成,促使肿瘤细胞凋亡。Objective To investigate the inhibitive effect human neuroblastoma in nude mouse models. Methods of antisense VEGF165 cDNA on growth and angiogensis of Three kinds of SH-SY5Y cells which had been stably transfected by sense VEGF165 cDNA, antisense VEGF165 cDNA and empty vector respectively, were inoculated subcutaneously to nude mice. The size of subcutaneous tumors was measured, and the morphology of tumor tissue was observed by microscope. The microvessel density （MVD） in tumor mass was analyzed by CD31 immunohistochemistry staining. Apoptotic cells were detected by the electron microscope. Results The growth of tumor mass, which was inoculated with antisense VEGF, significantly decreased in nude mice compared with empty vector group （P 〈0.05）, while the growth rate of tumor in the sense VEGF group significantly increased in nude mice （P 〈 0.05 ）. The morphologic investigation of SH-SY5Y cells in antisense group showed blood vessel sparsity, smaller nuclei, fewer cytoplasm, lesser mitotic figure compared with the empty vector group. The electron micrographs of SH-SY5Y cells in antisense group showed the characteristics of apoptosis： the chromatin condensed in masses at the periphery of nuclei, and the nuclear fragmentation were seen, but they were not seen in sense group and empty vector group. MVD in rumor mass of the antisense group significantly decreased as compared with empty vector （ P 〈 0. 01 ）, and MVD of sense group significantly increased （ P 〈 0.01 ）. Conclusion Antisense VEGF165 cDNA transfection inhibits the proliferation SH-SY5Y cells in the nude mice, decrease angiogensis and increase tumor cells apoptosis.

关 键 词：血管内皮生长因子 反义 神经母细胞瘤 微血管密度 超微结构 

分 类 号：R739.4[医药卫生—肿瘤]