| | | 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
| | 护理医学论文 作者：张吉成，陈燕凌，张德新，陈宏明，杨定忠，王作仁
【关键词】 胆囊肿瘤；血管抑素；抗血管生成；基因转染
Experimental study of antiangiogenic therapy with mouse angiostatin gene transduction in gallbladder cancer
【Abstract】 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 GBCSD 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 ECV304 was observed in vitro. The nude mice were divided into 2 groups: angiostatin group and control group implanted with GBCSD/pcDNA3.1(+)angiostatin cells and GBCSD 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 GBCSD/pcDNA3.1(+)angiostatin cells. The angiostatin protein could inhibit the proliferation of vascular endothelial cells ECV304 dependent on bFGF (P<0.01). In addition, the nude mice experiment showed that tumorigenic capability of the GBCSD/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.
【Keywords】 gallbladder neoplasms; angiostatin; antiangiogenesis; gene transfection
【摘要】 目的护理医学论文：研究重组小鼠血管抑素基因真核表达质粒转染的护理医学论文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GBCSD，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.
【关键词】 胆囊肿瘤；血管抑素；抗血管生成；基因转染
0引言
胆囊癌预后极差,故积极探索胆囊癌的基因治疗具有非常重要的意义.肿瘤的生长依赖于丰富的血供,通过抑制肿瘤新生血管形成就可以使肿瘤缺血坏死达到治疗目的.血管抑素(angiostatin)是1994年OReilly等［1］发现的能强烈抑制血管内皮细胞生长和新生血管形成的蛋白质.为此，我们拟应用重组小鼠血管抑素基因的真核表达质粒转染胆囊癌细胞株, 检测血管抑素的蛋白表达，并观察其对血管内皮细胞的作用和对裸鼠种植性胆囊癌生长的影响.探索血管抑素在胆囊癌血管抑制基因治疗中的意义.
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