Gene transfer of kringle 1-5 suppresses tumor development and improves prognosis of mice with hepatocellular carcinoma
Gene transfer of kringle 1-5 suppresses tumor development and improves prognosis of mice with hepatocellular carcinoma
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DOI：
10.1053/j.gastro.2006.02.020
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发表时间：
2006-04-01
期刊：
影响因子：
29.4
通讯作者：
Cao, YH
中科院分区：
文献类型：
--
作者：
Torimura, T;Ueno, T;Cao, YH
背景和目的：最近的研究表明 kringle 1-5 具有有效且特异性的抗血管生成活性。在这里，我们研究了kringle 1-5基因转移对小鼠肝细胞癌的抗肿瘤作用。方法：通过四唑检测评估kringle 1-5蛋白对牛毛细血管内皮细胞增殖的抑制作用。为了研究肿瘤生长、肝内转移和存活，将脂质体/kringle 1-5互补DNA复合物静脉注射到预先植入肝脏中的3种肝癌细胞系中的一种的裸鼠中。通过免疫组织化学和蛋白质印迹测试了 kringle 1-5 的产生。量化瘤内血管密度。通过蛋白质印迹法检测肿瘤中血管内皮生长因子、血管生成素-1和血管生成素-2的表达。测量血清丙氨酸转氨酶和甲胎蛋白水平以及体重。结果：纯化的kringle 1-5以剂量依赖性方式抑制牛毛细血管内皮细胞的增殖。 kringle 1-5的基因转移导致血管密度显着降低，抑制了3种肝癌细胞系的肿瘤生长和血清甲胎蛋白水平，延长了生存期，并减少了肝内转移的数量。在分析的血管生成因子中，kringle 1-5 降低了血管生成素-2 的表达水平。在肝癌细胞和肝脏肝细胞中检测到kringle 1-5蛋白的表达。然而，它并没有改变血清丙氨酸氨基转移酶水平和体重，这表明 kringle 1-5 没有严重的副作用。结论：使用 kringle 1-5 互补 DNA 进行抗血管生成基因治疗是一种有前景的安全有效的抑制肝细胞癌生长的策略。
Background & Aims: Recent studies indicate that kringle 1-5 has a potent and specific antiangiogenic activity. Here, we investigated the antitumor effect of kringle 1-5 gene transfer on hepatocellular carcinoma in mice. Methods: The inhibitory effect of kringle 1-5 protein on proliferation of bovine capillary endothelial cells was evaluated by a tetrazolium-based assay. To study tumor growth, intrahepatic metastasis, and survival, liposome/kringle 1-5 complementary DNA complexes were injected intravenously in nude mice preimplanted with :1 of 3 hepatoma cell lines into the liver. Production of kringle 1-5 was tested by immunohistochemistry and Western blotting. Intratumoral vessel density was quantified. Expression of vascular endothelial growth factor, angiopoietin-1, and angiopoietin-2 in tumors was examined by Western blotting. Serum alanine aminotransferase and alpha-fetoprotein levels and body weights were measured. Results: Proliferation of bovine capillary endothelial cells was inhibited by purified kringle 1-5 in a dose-dependent manner. Gene transfer of kringle 1-5 caused a significant reduction in vessel density with suppression of tumor growth of the 3 hepatoma cell lines and serum alpha-fetoprotein levels, prolonged the survival period, and reduced the number of intrahepatic metastases. Among the analyzed angiogenic factors, kringle 1-5 reduced angiopoietin-2 expression levels. Expression of kringle 1-5 protein was detected on hepatoma cells and hepatocytes in the liver. However, it did not alter serum alanine aminotransferase levels and body weights, suggesting kringle 1-5 lacks severe side effects. Conclusions: Antiangiogenic gene therapy with kringle 1-5 complementary DNA is a promising safe and effective strategy for suppression of growth of hepatocellular carcinoma.