NRP -1 b1/b2单克隆抗体对裸鼠胃癌移植瘤生长的影响

目的：探讨神经纤毛蛋白1（neuropilin -1，NRP -1）在胃癌细胞 BGC823中的表达情况，并观察自主研发的抗 NRP -1 b1/ b2单克隆抗体（NRP -1 mAb）对裸鼠胃癌移植瘤生长的影响。方法：基于实验室前期已成功制备出纯度和浓度均较高的 NRP -1 mAb。利用流式细胞术检测该抗体亲和性、细胞免疫荧光定位检测 NRP -1的分布及 NRP -1 mAb 特异性。建立胃癌移植瘤裸鼠模型，向移植瘤中注射不同浓度的 NRP -1 mAb，研究抗体对胃癌裸鼠皮下移植瘤生长的影响。2周后根据瘤体大小得出抑瘤率。最后免疫组化分析癌组织内血管内皮生长因子（VEGF）的表达情况。结果：胃癌细胞 BGC -823上存在 NRP -1，且主要分布在细胞膜上，能与自制的 NRP -1 mAb 有效结合。通过动物实验，NRP -1 mAb 能有效抑制移植瘤的生长，高浓度抗体较低浓度对移植瘤抑制作用更明显，NRP -1 mAb 治疗组中 VEGF 表达较空白组低（P <0.01）。结论：胃癌细胞 BGC -823细胞膜上表达 NRP -1，NRP -1 mAb 能特异性结合胃癌细胞上 NRP -1蛋白，NRP -1 mAb 能抑制胃癌裸鼠移植瘤的生长，且与浓度呈正相关，可能与下调 VEGF 表达有关。

The effects of NRP - 1 b1/b2 monoclonal antibody on the growth of stomach transplan-ted tumor in nude mouse

Objective:Explore neuropilin - 1(NRP - 1)expression in gastric cancer BGC823 cells,and observe the effects of NRP - 1 mAb which was successfully independed produced on the growth of stomach transplanted tumor in nude mouse. Methods:Our laboratory had successfully produced NRP - 1 monoclonal antibody(NRP - 1 mAb) with high purity and concentration on the early stage. The affinity and specificity of NRP - 1 mAb and the location de-tection of NRP - 1 were detected by flow cytometry and cell immunofluorescence techniques. Different concentration of NRP - 1 mAb were injected into the transplantation tumor of BGC - 823 xenografts nude mice,then observed the rate of tumor inhibition after two weeks. Using immunohistochemistry tested the expression of vascular endothelial growth factor(VEGF). Results:The NRP - 1 was especially situated on the membrane of gastric cancer cell lines in BGC -823 cells. In animal experiments,NRP - 1 mAb can effectively inhibit the growth of the transplanted tumor. The more obvious inhibitory effect was observed in the high concentration group. VEGF expression of NRP - 1 mAb in the treat-ment group was lower than those of blank group(P < 0. 01). Conclusion:NRP - 1 express on the cell membrane of gastric cancer cells BGC - 823. NRP - 1 mAb can combine with NRP - 1 protein in gastric cancer cells. NRP - 1 mAb can inhibit the growth of the transplanted tumor,and was positively correlated with concentration. This may be phase cooperation with VEGF.