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

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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.