Basic research of gene therapy for cancer utilizing vascular endothelial cells.

利用血管内皮细胞进行癌症基因治疗的基础研究。

• 批准号: 06807095
• 负责人: NAKAMURA Hiroshige
• 金额: $ 1.22万
• 依托单位: Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06807095/
• 关键词: Gene Therapy; Endothelial Cells; Vein; Lipofectin; IFN gamma; Cancer; 遺伝子導入; IFN; 培養

The purpose of this project is to activate anti-tumor immunity according to transfecting some kinds of cytokine genes, Interferon or TNF,into vascular endothelial cells of feeding artery within tumor tissue. First of all, we try to transfect lac Z gene into human endothelial cells of great saphenous vein which was corrected by the operation for primary varicose vein. Endothelial cells was cultured in Medium 199 with 20% heat inactivated patients' serum. Using lipofection method, 20 mu g Lac Z gene was tranfected into 5*10^5 endothelial cells. Lac Z activity was observed in 10% of endothelial cells. Next protocol was to trasfect IFN expression vector, pMNMS-IFN,into human endothelial cells. Same as first protocol, INF activity was observed 10% of endothelial cells and its concentration of supernatant was very low level, 1000pg/l*10^7/ml. Finally, we attempt to transfect IFN gene into human vein. But few trasfected cells was observed within venous tissue. We conclude that it is very important to increase efficiency of transfection in order to give some reality to this gene therapy.

该项目的目的是通过将某些细胞因子基因、干扰素或TNF转染到肿瘤组织内供血动脉的血管内皮细胞中来激活抗肿瘤免疫。首先，我们尝试将lac Z基因转染到经过原发性静脉曲张手术矫正的人大隐静脉内皮细胞中。内皮细胞在含有20%热灭活患者血清的培养基199中培养。采用脂转染法，将20μg Lac Z基因转染至5*10^5的内皮细胞中。在 10% 的内皮细胞中观察到 Lac Z 活性。下一步方案是将 IFN 表达载体 pMNMS-IFN 转染至人内皮细胞中。与第一个方案相同，观察到10%的内皮细胞有INF活性，并且其上清液浓度非常低，为1000pg/l*10^7/ml。最后，我们尝试将IFN基因转染到人静脉中。但在静脉组织内观察到很少的转染细胞。我们的结论是，为了使这种基因疗法具有一定的现实意义，提高转染效率非常重要。