Combination cancer gene therapy in endostatin and suicide gene for metastatic murine mammary cancers

内皮抑素和自杀基因联合治疗转移性小鼠乳腺癌

• 批准号: 15591368
• 负责人: SHIBATA Masa-aki
• 金额: $ 2.37万
• 依托单位: Osaka Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591368/
• 关键词: Angiogenesis; Endostatin; Angiostatin; Mammary carcinoma; Cancer gene therapy; Electrogene transfer; Mouse; Human umbilical vascular endothelial cell; Mammry carcinoma; Elrctrogene transfer

Experimental mammary cancer therapy in mice was conducted with electrogene transfer of a non-viral plasmid vector containing angiostatic genes. In vitro study : HUVECs transfected with endostatin (pEndo), angiostatin (pAngio) and both fusion genes (pEndo:pAngio) were inhibited the tubular formation on Matrigels. In vivo study : Experiment 1. Mammary tumors induced by inoculation of BALB/c mice with BJMC3879 cells were subsequently treated by direct injection of pEndo,pAngio,pEndo:pAngio or control vector once a week for 7 weeks. Significantly reduced tumor volumes were observed for the pEndo or pEndo:pAngio groups in experimental week 1 and thereafter throughout the study. In the pAngio group, reduced tumor volume was only observed in week 7. Experiment 2. Using the same mammary cancer model, combination therapy in pEndo with suicide gene (HSVtk/GCV) was conducted. Tumor volumes were significantly suppressed in the pEndo,pHSVtk/GGCV and the combination groups. In the combination group, the tumor volumes were further suppressed as compared with respective alone treatment groups. In therapeutic groups, metastases to lungs or lymph nodes were also significantly decreased. However, summative or synergistic effects were not observed in the combination group on metastases. Microvessel density within tumor tissues was significantly decreased in the pEndo or the combination groups. Decreased DNA synthesis and elevated apoptosis were also found in all therapeutic groups. in addition, numbers of lymphatic vessels with migrating cancer cells in their lumens were significantly inhibited in pEndo or the combination groups. We therefore conclude that endostatin gene with in vivo electrogene transfer can result in suppression of mammary tumor growth and metastases. In addition, endostatin in combination therapy with HSVtk/GCV further suppressed tumor growth but summative effects were not observed on reduction of metastases.

通过含有血管抑制基因的非病毒质粒载体的电基因转移对小鼠进行实验性乳腺癌治疗。体外研究：用内皮抑素（pEndo）、血管抑制素（pAngio）和两种融合基因（pEndo：pAngio）转染的HUVEC在基质胶上抑制管状形成。体内研究： 实验1.通过用BJMC3879细胞接种BALB/c小鼠诱导的乳腺肿瘤随后通过每周一次直接注射pEndo、pAngio、pEndo:pAngio或对照载体进行治疗，持续7周。在实验第一周以及此后的整个研究过程中，观察到 pEndo 或 pEndo:pAngio 组的肿瘤体积显着减小。在pAngio组中，仅在第7周时观察到肿瘤体积缩小。 实验2.使用相同的乳腺癌模型，进行pEndo与自杀基因(HSVtk/GCV)的联合治疗。 pEndo、pHSVtk/GGCV 和联合组的肿瘤体积被显着抑制。在联合组中，与各个单独治疗组相比，肿瘤体积被进一步抑制。在治疗组中，肺部或淋巴结的转移也显着减少。然而，在联合组中没有观察到对转移的总结性或协同效应。 pEndo 组或联合组中肿瘤组织内的微血管密度显着降低。在所有治疗组中还发现 DNA 合成减少和细胞凋亡增加。此外，在pEndo组或联合组中，管腔内具有迁移癌细胞的淋巴管数量显着受到抑制。因此，我们得出结论，内皮抑素基因与体内电基因转移可以抑制乳腺肿瘤的生长和转移。此外，内皮抑素与HSVtk/GCV联合治疗进一步抑制了肿瘤生长，但没有观察到对减少转移的累积效应。