重组小鼠血管抑素基因转染抗血管生成治疗胆囊癌的实验研究  

Experimental study of antiangiogenic therapy with mouse angiostatin gene transduction in gallbladder cancer

作　　者：张吉成[1] 陈燕凌[1] 张德新[2] 陈宏明[1] 杨定忠[3] 王作仁[3] 

机构地区：[1]福建医科大学附属协和医院肝胆外科,福建福州350001 [2]第四军医大学西京医院全军消化病研究所,陕西西安710033 [3]西安交通大学第一医院肝胆外科,陕西西安710061

出　　处：《第四军医大学学报》2007年第5期461-464,共4页Journal of the Fourth Military Medical University

基　　金：福建省自然科学基金(C0540008)

摘　　要：目的:研究重组小鼠血管抑素基因真核表达质粒转染的胆囊癌细胞表达具有抑制血管内皮细胞生长活性的血管抑素蛋白,以及对裸鼠种植性胆囊癌生长的抑制作用.方法:应用脂质体lipofectamine2000将重组小鼠血管抑素基因的真核表达质粒pcDNA3.1(+)-angiostatin转染胆囊癌细胞株GBC-SD,G418抗性筛选;以Western blot检测血管抑素的表达,以血管内皮细胞增殖分析检测其生物学活性,接种裸鼠并比较肿瘤体积和肿瘤微血管密度(MVD).结果:得到表达血管抑素的胆囊癌细胞克隆,其培养上清能有效抑制bFGF刺激的血管内皮细胞增殖(P<0.01);血管抑素组裸鼠肿瘤体积小于野生细胞对照组(P<0.01),免疫组化检测显示血管抑素组裸鼠肿瘤微血管密度(MVD)低于野生细胞对照组(P<0.05).结论:血管抑素基因转染对裸鼠种植性胆囊癌的生长有抑制作用,这是血管抑素组肿瘤组织内新生血管形成受到抑制而减少所致.AIM： To investigate the expression of mouse angiostatin gene transfected into gallbladder cancer cells, antiangiogenic activity of angiostatin protein and inhibitory effect of angiostatin on implanted gallbladder carcinoma of nude mice. METHODS： The recombinant vector pcDNA3.1 （ ＋ ）-angiostatin was transfected into gallbladder cancer cells GBC-SD with liposome lipofectamine2000. Angiostatin protein expression was examined by Western blot in the stable cell lines by G418 and its inhibitory effect on the vascular endothelial cells ECV-304 was observed in vitro. The nude mice were divided into 2 groups： angiostatin group and control group implanted with GBC-SD/ pcDNA3.1 （ ＋ ） -angiostatin cells and GBC-SD cells respectively. The carcinoma volume and microvessel density （MVD） of each group were compared and analyzed. RESULTS： After 30 d of transfection and selection with G418, macroscopic resistant cell clones were formed. Western blot showed that angiostatin protein was expressed and secreted by GBC-SD/pcDNA3. 1 （ ＋ ）- angiostatin cells. The angiostatin protein could inhibit the proliferation of vascular endothelial cells ECV-304 dependent on bFGF （P 〈0.01 ）. In addition, the nude mice experiment showed that tumorigenic capability of the GBC-SD/pcDNA3. 1 （ ＋ ）-angiostatin cells had been reduced significantly （ P 〈 0.01 ）. Immunochemistry study demonstrated that MVD of the angiostatin group was lower than that of the control group （ P 〈 0.05 ）. CONCLUSION： Angiostatin gene transduction may have potential value in the treatment of gallbladder cancer in the future.

关 键 词：胆囊肿瘤 血管抑素 抗血管生成 基因转染 

分 类 号：R735.8[医药卫生—肿瘤]