NRP-1单克隆抗体对乳腺癌裸鼠移植瘤生长的影响

目的探讨NRP-1单克隆抗体(NRP-1 MAb)的特异性,以及不同剂量的NRP-1 MAb治疗乳腺癌裸鼠移植瘤的疗效。方法 Western blot和共聚焦免疫荧光法检测NRP-1 MAb是否识别MCF7细胞上NRP-1蛋白。将MCF7细胞接种于BALB/c裸鼠皮下建立乳腺癌细胞移植瘤模型,并进行瘤组织传代。传代的肿瘤体积生长至300～500 mm^3时,随机分为对照组、NRP-1 MAb低剂量组、中剂量组和高剂量组,每组6只,给药7次。观察荷瘤裸鼠一般状况,测量瘤体大小及裸鼠体重。实验结束时剥离瘤体称重,提取组织蛋白,Western blot检测组织中VEGF蛋白和NRP-1蛋白的表达量。结果 NRP-1MAb成功识别MCF7细胞上的NRP-1蛋白;NRP-1 MAb能够有效抑制MCF7细胞裸鼠移植瘤的生长,低剂量组(1 mg/kg)抑瘤率为47.01%,中剂量组(5 mg/kg)抑瘤率为65.70%,高剂量组(10 mg/kg)抑瘤率为69.19%。结论 NRP-1 MAb能够识别并有效结合MCF7细胞膜上的NRP-1蛋白,且可抑制MCF7细胞移植瘤的生长,NRP-1 MAb抑制移植瘤的增长可能与下调NRP-1和VEGF表达有关。

Influence of NRP-1 Monoclonal Antibody on Growth of Breast Cancer Xenografts in Nude Mouse

Objective To investigate the specificity of NRP-1 monoclonal antibody (NRP-1 MAb), and the antitumor effect of NRP-1 MAb at the different doses on nude mice bearing breast cancer xenografts. Methods The recognition of NRP-1 protein in MCF7 cells by NRP-1 MAb were detected by Western blot and confocal immunofluorescent analysis. BALB/c nude mice were adopted to establish a tumor-bearing model by inoculating subcutaneously MCF7 cells, and then the passage of tumor tissues was carried out. When the tumor grew to 300-500 mm3, the xenografts nude mice were randomly divided into control group, low-dose group, middle-dose group and high-dose group, 6 mice in each group, and 7 doses in full course by subcutaneous injection. Then we observed the general status, tumor size and body weight in the routine. When the trial ended, relevant tumors were striped and weighed, and then we extracted the protein. Western blot was performed to examine the expression of VEGF and NRP-1. Results NRP-1 MAb could bind the NRP-1 protein in MCF7 cells, and significantly inhibit the growth of breast cancer xenograft in nude mice. The corresponding inhibition ratio reached 47.01%, 65.70% and 69.19% in low-dose group, middle-dose group and high-dose group, respectively. Conclusion NRP-1 MAb could recognize and bind NRP-1 on the membrane of MCF7 cells and inhibit the growth of the transplanted tumor. The downregulation of VEGF and NRP-1 may play an active role in tumor growth inhibition.