可溶性VEGF受体基因sflt-1与反义VEGF核苷酸对新生血管形成的影响

背景与目的:肿瘤组织中新生血管提供的大量营养物质和生长因子是肿瘤快速生长的关键,因此如何抑制肿瘤组织中新生血管的形成、促使肿瘤组织坏死也是肿瘤治疗中的一条值得探讨的途径。本文拟研究和观察可溶性血管内皮细胞生长因子(vascularendothelialgrowthfactor,VEGF)受体基因sflt-1与反义VEGF对新生血管形成的影响。方法:用Ad-反义VEGF感染肝癌细胞株MM45T.Li后,观察反义VEGF对MM45T.Li分泌VEGF的影响;将人工重组的3'ΔFlt-1蛋白(C末端缺失的Flt-1蛋白)加入条件培养液中,观察3'ΔFlt-1对VEGF刺激的脐静脉血管内皮细胞(humanumbilicalvascularendotheliumcell,HUVEC)增殖的影响;并将Ad-反义VEGF、Ad-sflt-1注射于鸡胚尿囊绒毛膜中,观察sflt-1、反义VEGF对鸡胚新生血管形成的影响。结果:反义VEGF重组腺病毒感染MM45T.Li细胞后,细胞培养上清中VEGF浓度仅为对照组中VEGF浓度的15%(P<0.01);在含有3'ΔFlt-1蛋白的调价培养液中,HUVEC的增殖明显减慢,在一定的范围内与剂量呈负相关;两者都能有效地抑制鸡胚新生血管的形成。结论:sflt-1与反义VEGF均能有效抑制新生血管的形成,两者联合能增强抑制效果,但其作用机制不同。

Effect of soluble vascular endothelial growth factor (VEGF) receptor gene (sflt-1) and antisense VEGF nucleotide on neovascularization

BACKGROUND & OBJECTIVES: Nutrients and growth factor provided by new blood vessel in tumor tissue is critical for fast growth of tumor, therefore, how to inhibit neovascularization and speedup neocrosis of tumor tissue is a promising way to treat tumor. This article emphasizes on both the blockage effect of soluble VEGF receptor gene (sflt-1) and antisense VEGF nucleotide on VEGF in tumor tissue, in order to inhibit its neovascularization. METHODS: Effect of antisense VEGF on VEGF secretion in MM45T. Li cells was observed after infection with Ad-antisense VEGF; Effect of recombinant soluble VEGF receptor protein (3' delta Flt-1) on proliferation of (human umbilical vascular endothelial cell) HUVEC induced by VEGF was investigated; Effect of both antisense VEGF and sflt-1 on neovascularization were tested after Ad-anti VEGF or Ad-sflt-1 injected in chorion of chicken embryo. RESULTS: VEGF concentration from infected MM45T. Li by recombinant Ad-antisense VEGF is only 15% of that from control group (P < 0.01); the proliferation of HUVEC induced by VEGF in condition medium containing 3' delta Flt-1 is significantly reduced, and negatively related with dose within certain range; Both sflt-1 and antisense VEGF nucleotide obviously inhibited formation of new blood vessel, even caused dead embryo. CONCLUSION: Both sflt-1 and antisense VEGF could effectively inhibited neovascularization separately, combination therapy can enhance the inhibitory effect, but the mechanism is different.