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

本课题的目的是通过将干扰素（Interferon）或肿瘤坏死因子（TNF）等多种细胞因子基因转染到肿瘤组织内供血动脉的血管内皮细胞中，激活抗肿瘤免疫。首先，我们尝试将lac Z基因导入经手术矫正的人大隐静脉内皮细胞。内皮细胞在含有20%热灭活患者血清的Medium 199中培养。采用脂质体转染法，将20 μ g Lac Z基因转染5 × 10^5个内皮细胞。在10%的内皮细胞中观察到Lac Z活性。下一步是将IFN表达载体pMNMS-IFN转染人内皮细胞。与第一方案相同，10%的内皮细胞观察到INF活性，其上清液浓度非常低，1000 pg/l*10^7/ml。最后，我们尝试将IFN基因导入人静脉。静脉组织中未见转染细胞。我们的结论是，这是非常重要的，以提高转染效率，以使一些现实的基因治疗。

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.