反应停的抗新生血管形成及抗肿瘤作用研究

Study on antiangiogenesis and antitumor of thalidomide

目的 研究反应停的抗新生血管形成及抗肿瘤作用。方法 在HUVECs中MTT法检测细胞活性,电镜观察细胞死亡类型,流式细胞仪定量细胞凋亡率;用鸡胚尿囊膜模型观察新生血管形成;肉瘤S180小鼠体内模型观察反应停的抗肿瘤作用,并通过免疫组化观察反应停对肿瘤组织微血管数的影响。结果 反应停对HUVECs有直接抑制作用,IC50为(22 .91±1. 74) μmol·L-1;经反应停作用48h后的HU VECs,随剂量增加,电镜下可见核不规则,染色体浓集,空泡状内质网及凋亡小体等不同时期细胞凋亡现象;流式细胞检测显示反应停可依剂量诱导HUVECs凋亡或坏死;鸡胚尿囊膜新生血管形成模型中,反应停抑制新生血管形成的阳性率随剂量而增加;小鼠S180肿瘤移植模型中,反应停单独使用,虽可明显减少肿瘤血管密度, 无抑制肿瘤生长作用;与环磷酰胺联合使用,可减少环磷酰胺用量,与环磷酰胺有协同抗肿瘤作用。结论 反应停有抗新生血管形成作用,单独应用对小鼠S180肿瘤无抑制作用,但与环磷酰胺有协同作用。

Aim To study antiangiogenesis and antitumor of thalidomide.Methods In HUVECs, cell viability was determined by MTT assay;death type was observed by electron microscope; ratio of apoptosis was quantitated by flow cytometry.Angiogenesis was tested in chicken embryo chorioallantoic membrane.Effect of thalidomide on S_(180) was examined in homograft mice and microvascular counts were detected through immunochemical staining method.Results Thalidomide might inhibite the growth of HUVECs with a IC_(50) value of(22.91±1.74) μmol·L^(-1),cells treated by thalidomide for 48 h displayed morphological characteristics of different stages associated with apoptosis,which were irregular nucleus, condensed chromatin, ballooning endoplasmic reticulum, apoptotic bodies,under electron microscope.Thalidomide might be able to cause apoptosis or necrosis of HUVECs in flow cytometry and raised positive of antiangiogenesis with increasing of dosage in chicken embryo chorioallantoic membrane. Thalidomide as a single agent might not significantly prevent tumor growth but decrease microvascular counts in tumors, however, in combination with cyclophosphamide, thalidomide could decrease dosage of cyclophosphamide and enhance antitumor of cyclophosphamide.Conclusion Thalidomide might hold back angiogenesis,as a single agent, could not significantly prevent S_(180) tumor from growing,but acted synergistically with cyclophosphamide.