内皮抑素基因抑制兔VX_2肝癌肝动脉栓塞后肿瘤血管生成的实验研究  

Endostatin gene therapy suppresses the angiogenesis of rabbit VX_2 liver tumor after transcatheter arterial embolization

作　　者：廖晓峰[1] 孙华朋[1] 易继林[2] 

机构地区：[1]华中科技大学同济医学院附属襄樊中心医院普外科,湖北襄樊441021 [2]华中科技大学同济医学院附属同济医院普外科,湖北武汉430030

出　　处：《现代肿瘤医学》2009年第6期1021-1026,共6页Journal of Modern Oncology

摘　　要：目的:研究内皮抑素基因治疗对介入栓塞后兔VX2肝肿瘤血管生成的抑制作用。方法:建立兔VX2肝癌模型,分为栓塞(TAE)组,肝动脉注射(IHA)组和瘤内注射(IT)组。三组动物均经肝动脉注入超液化碘油进行肝动脉栓塞。IHA组在栓塞的同时,经肝动脉注入携带内皮抑素基因的真核表达质粒pCEP-Pu,IT组在栓塞的同时直接瘤内注射pCEP-Pu。测定栓塞前后血清谷丙转氨酶(ALT)含量。栓塞后第7天处死动物,计算抑瘤率。免疫组化方法检测肿瘤微血管密度(MVD)及血管内皮生长因子(VEGF)蛋白的表达,Western印迹杂交测定VEGF蛋白表达,逆转录聚合酶链反应(RT-PCR)检测VEGF mRNA的表达。结果:IHA组和IT组MVD分别为(16.5±5.7)和(18.8±7.4),显著低于TAE组MVD(28.6±10.6)(P<0.05)。组化及Western印迹杂交结果均显示IHA组和IT组VEGF蛋白表达显著低于TAE组。RT-PCR结果显示IHA组和IT组VEGF165 mRNA表达显著低于TAE组(P<0.01),分别为(2.03±0.42),(2.01±0.39)和(2.58±0.42)。三组栓塞前后血清ALT水平无差异(P>0.05)。三组栓塞后肿瘤体积差异无显著性(P>0.05)。结论:介入栓塞的同时经肝动脉注射内皮抑素基因可方便、有效地阻止栓塞后残留肿瘤的血管生成,无明显的肝脏毒性。Objective：To investigate the effectiveness of endostatin gene therapy for inhibition of rabbit VX2 liver tumor angiogenesis after transcatheter arterial embolization. Methods ： VX2 carcinoma was implanted in the left medial lobes of 30 New Zealand White rabbit livers. The animals were divided into three groups of ten rabbits each randomly. Fourteen days later,a silicon catheter was inserted into the left hepatic artery of rabbit with VX2 hepatic tumor and infusion was performed via the hepatic artery using Lipiodol in all experiment animals. Meanwhile, the plasmid pCEP - Pu expressing endostatin was delivered accompanied with Lipiodol via the hepatic artery in IHA group and injected directly by intratumoral administration in IT group. Before and after embolization, blood serum alanine transaminase （ALT） was measured. Rabbits were sacrificed 7 days after infusion and the volume of tumors was evaluated. CD31 was immunohistochemically used to count microvessel density （MVD） in tumor. Immunohistochemistry and western blotting were used to detect expression of vascular endothelial growth factor （VEGF） protein, and reverse transcription - polymerase chain reaction （RT- PCR） was used to investigate VEGF mRNA in tumor. Results： MVD of IHA group and IT group were （16.5 ± 5.7 ） and （18.8 ± 7.4）, respectively. Both of them were lower than that of TAE group （28.6 ± 10.6） （P 〈0.05）. Expressions of VEGF protein were reduced in both IHA group and IT group, as compared with the TAE group （P 〈 0.05 ）. In mRNA level, VEGF165 mRNA was significantly lower in IHA group （2.03 ±0.42） and IT group （2.01 ±0.39） than TAE group （2.58 ±0.42） （P〈0.01）.There were no significant differences of the average tumor sizes and the content of serum ALT among the three groups. Conclusion： Gene therapy with expression plasmid carrying endostatin delivered by intrahepatic artery route can inhibit the angiogenesis of rabbit VX2 liver tumor undergoing transeatheter arterial embolizati

关 键 词：基因治疗 内皮抑素 抗血管新生 肝肿瘤 化学栓塞 

分 类 号：R735.7[医药卫生—肿瘤] R734.2[医药卫生—临床医学]