重组可溶性KDR及其抗体对内皮细胞的增殖抑制作用  

The Inhibitory Effect of Recombinant Solube KDR and Its Antibody on Endotheliocyte Proliferation

作　　者：刘丽[1] 肖畅[1] 王烨[1] 曹颖[1] 

机构地区：[1]中国人民解放军空军总医院空军临床分子生物学中心实验室,北京100036

出　　处：《中国肿瘤生物治疗杂志》2002年第1期23-26,共4页Chinese Journal of Cancer Biotherapy

摘　　要：目的 :分析可溶性KDR(sKDR)及其抗体对内皮细胞增殖的抑制作用。方法 :通过ELISA分析大肠杆菌表达的sKDR纯化产物与VEGF16 5结合的能力 ;sKDR免疫家兔制备KDR2 6 2抗血清 ,Wensternblot分析该抗血清与KDR2 6 2蛋白结合的特异性 ;用3 H TdR掺入法 ,MTT法和细胞计数 3种方法分析重组sKDR及其抗体对内皮细胞的增殖抑制作用。结果 :sKDR蛋白可与VEGF16 5特异性结合 ,肝素可增强其结合能力。KDR2 6 2抗血清具有特异性识别KDR蛋白的能力 ,其滴度为 1∶2 0 0 0。用3 H TdR掺入法和MTT 2种方法分析sKDR蛋白及其抗体对内皮细胞的增殖抑制作用结果一致 ,sKDR蛋白浓度在 10 μg/ml,2μg/ml,0 .4μg/ml时对内皮细胞增殖抑制率平均为 5 6 %,44 %和 32 %;抗KDR2 6 2抗体在稀释度为 5 0 ,2 0 0和 80 0倍时对内皮细胞增殖的抑制率平均为 70 %,5 6 %和 43%。细胞计数法测定结果 ,sKDR及抗KDR2 6 2抗体组与VEGF16 5单独刺激组 ,GST和PBS对照组相比 ,内皮细胞增殖被明显抑制 ,随浓度增加抑制活性明显增强 ,但抗体的抑制活性比sKDR蛋白高。结论 :大肠杆菌表达的sKDR及其抗体对内皮细胞均具有明显的增殖抑制作用。Objective: To investigate the inhibitory effect of soluble KDR(sKDR) and its antibody on endotheliocyte(EC) proliferation.Methods: The binding of sKDR with VEGF was detected by ELISA. The KDR262 antiserum was prepared from rabbit and the specificity was defined by Western blot. The inhibition of EC proliferation was analyzed by 3H-TdR up-take, MTT and cell counter. Results: The sKDR could bind to VEGF165 specially, the ability was enhanced by heparin. The specific binding of KDR262 antiserum to KDR was also detected. The inhibitory rate of sKDR(10,2,0 4μg/ml) on EC proliferation was 56%,44%,32% respectively. The inhibitory rate of KDR262 antiserum (1∶50,1∶200,1∶800) was 70%, 56%,43% respectively. The cell counter also showed that EC proliferation was inhibited significantly in sKDR group and KDR262 antibody group, vs VEGF165 group, GST group and PBS group. The inhibitory effect was characterized by concentration-dependent and was higher in KDR262 antibody group than that in sKDR group. Conclusion: sKDR expressed by E.coli and its antibody had significant inhibitory influence on EC proliferation.

关 键 词：重组可溶性KDR 抗体 内皮细胞 抑制作用 血管内皮细胞生长因子 抗血管形成 细胞增殖 

分 类 号：R730.231[医药卫生—肿瘤]