羟喜树碱联合热疗对体内外血管生成的抑制作用

Effect of hydroxycamptothecin with thermotherapy on angiogenesis in vivo and in vitro

目的:探讨羟喜树碱联合应用热疗(thermotherapy)对体内外血管生成的抑制作用.方法:采用MTT法观察羟喜树碱联合热疗对人微血管内皮细胞(HMVEC)、人肝癌细胞HepGⅡ增殖的影响.利用三维血管模型观察羟基喜树碱联合热疗对HMVEC迁移的影响.利用鸡胚绒毛尿囊膜(CAM)模型,观察羟喜树碱对鸡胚绒毛尿囊膜血管生成的影响.结果:羟喜树碱在20～40 ng/ml、5～80 ng/ml浓度范围内,具有抑制血管内皮细胞增殖和迁移作用,内皮细胞存活率(68.2%～44.7%),均与浓度呈负相关(相关系数r分别为-0.906、-0.469,P值分别为0 000 03、0.005 1);具有抑制鸡胚绒毛尿囊膜血管生成的作用;而在5～80ng/ml浓度范围内,对人肝癌细胞无细胞毒作用(细胞存活率:100%～90%).20 ng/ml的羟喜树碱与热疗联合应用在体外具有抗血管生成的协同效应;40 ng/ml的羟喜树碱与热疗联合应用在体外具有抗血管生成的次加效应.结论:小剂量羟喜树碱(20～40 ng/ml)联合热疗在体内外均具有抑制血管增殖和迁移协同或次加作用.

Purpose： To study the effects of hydroxyeamptothecin with thermotherapy on anti-angiogenesis in vitro and in vivo. Methods：We chose human microvascular endothelial cell （HMVEC） cuhure and chick embryo ehoriallantoic membrane （CAM） model and used MTF and re,tuber-calculating methods to observe hydroxycamptotheeine on HMVEC＇s proliferation , sprouts and CAM blood vessels＇ formation. Results： The survival rate of endothelial cells was in the range of 68.2%-44.7% within the dose of 20-40 ng/ml and 5-80 ng/ml and was negatively correlated with the concentration （ correlation coefficient was - 0. 906, -0. 469. P = 0. 00003.0. 0051 ）. Hydroxycamptotheeine could significantly suppress the endothelial cells＇ proliferation and the sprouts. Hydruxycamptotheciue could significantly suppress CAM vessels. The survival rate of HepG 11 cells is in the range 100% -90% within the dose of 5-80 ng/ml. There was no cytotoxicity. There was a syuergestic anti-angiogenetic effect when hydroxycamptothecin （20 ng/ml） was combined with thermotherapy in vitro while there was additive effect when hydroxycamptothecin （40 ng/ml） was combined with thermotherapy in vitro. Conclusions： This experiment shows that small doses of hydroxycamptothecine （20-40 ng/ml） with thermotherapy has anti-angiogenetic synergestic or additive effect on proliferation and migration both in vivo and in vitro.