靶向NRP1单克隆抗体对黑色素瘤细胞B16F10的增殖、迁移和凋亡的影响

发布时间：2018-07-10 02:27

本文选题：神经菌毛素单克隆抗体 + 黑色素瘤细胞　；参考：《中国药学杂志》2014年20期

【摘要】：目的研究靶向神经菌毛素1(neuropilin1,NRP1)单克隆抗体(NRP1 mAb)对黑色素瘤细胞B16F10增殖、迁移和凋亡的影响,并探讨其作用机制。方法 SDS-PAGE检测神经菌毛素1单克隆抗体的纯度;蛋白质印迹法、免疫荧光和免疫组化检测神经菌毛素1单克隆抗体的特异性和B16F10细胞及其移植瘤上神经菌毛素1的表达;四甲基偶氮唑蓝法、划痕实验、Annexin V-FITC/PI双标记流式细胞术等方法检测不同浓度神经菌毛素1单克隆抗体对黑色素瘤细胞增殖、迁移和凋亡的影响;蛋白质印迹法检测神经菌毛素1单克隆抗体作用后细胞内相关信号蛋白的表达。结果纯化的神经菌毛素1单克隆抗体具有高纯度和特异性,同时B16F10细胞及其移植瘤上神经菌毛素1高表达。神经菌毛素1单克隆抗体能有效抑制B16F10细胞的增殖和迁移,抑制作用呈时间和浓度依赖性(P0.05)。神经菌毛素1单克隆抗体能显著诱导B16F10细胞凋亡,并呈剂量依赖关系(P0.05)。抗体作用后B16F10细胞中磷酸化的Akt和ERK1/2表达下调,Bcl-2和Caspase 3蛋白表达下调,Bax蛋白的表达上调。结论纯化的神经菌毛素1单克隆抗体能有效地抑制B16F10增殖,迁移并明显诱导凋亡,其可能通过抑制膜上神经菌毛素1与相关细胞因子结合而减少其介导的PI3K/Akt和MEK/ERK1/2信号转导途径的活化来抑制细胞增殖和迁移,并通过Bcl-2/Bax/Caspase3途径诱导细胞凋亡,从而发挥其抗肿瘤作用。
[Abstract]:Objective to investigate the effects of neuropilin 1 (NRP1) monoclonal antibody (NRP1 mAb) on the proliferation, migration and apoptosis of melanoma cell line B16F10, and to explore its mechanism. Methods the purity of monoclonal antibody against neuromycin 1 was detected by SDS-PAGE, the specificity of monoclonal antibody against neuromycin 1 and the expression of neuromycin 1 on B16F10 cells and its transplanted tumor were detected by Western blot, immunofluorescence and immunohistochemistry. The effects of different concentrations of neuromycin-1 monoclonal antibody on the proliferation, migration and apoptosis of melanoma cells were detected by MTT assay and Annexin V-FITC / Pi double labeling flow cytometry. The expression of related signal proteins was detected by Western blot. Results the purified monoclonal antibody was of high purity and specificity, and the expression of neuromycin 1 was high in B16F10 cells and its transplanted tumor. Neuromycin 1 monoclonal antibody could effectively inhibit the proliferation and migration of B16F10 cells in a time-and concentration-dependent manner (P0.05). Neuromycin 1 monoclonal antibody could induce apoptosis of B16F10 cells in a dose-dependent manner (P0.05). The expression of phosphorylated Akt and ERK1 / 2 in B16F10 cells was down-regulated by antibodies, and the expression of Bcl-2 and caspase 3 was down-regulated in B16F10 cells. Conclusion the purified monoclonal antibody against neuromycin 1 can effectively inhibit the proliferation, migration and apoptosis of B16F10. It may inhibit cell proliferation and migration by inhibiting the activation of PI3K / Akt and MEK / ERK1 / 2 signal transduction pathways, and induce cell apoptosis through Bcl-2 / Bax-caspase3 pathway. In order to play its anti-tumor effect.
【作者单位】：厦门大学医学院抗癌研究中心;
【基金】：国家自然科学基金资助项目(30973485,81172970) 福建省属公益类科研专项经费(2011R1039-1,2013J0138)
【分类号】：R739.5

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