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	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; Effects of Anti-NRP1 Monoclonal Antibody on Cell Proliferation,Migration and Apoptosis of Melanoma B16F10 Cells
	吕莎 ; 李哲 ; 王显江 ; 王生育 ; 杨芸 ; 罗芳洪 ; 吴婷 ; 颜江华
	2014-10-22
关键词	神经菌毛素1单克隆抗体 黑色素瘤细胞 增殖 迁移 凋亡 NRP1 mAb melanoma proliferation migration apoptosis
英文摘要	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(P<0.05)。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(P<0.05)。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。; OBJECTIVE To investigate the effect of anti NRP1 monoclonal antibody( NRP1 mAb) on melanoma cell B16F10 proliferation,migration and apoptosis,and explore its mechanism.METHODS Western blot,immunofluorescent and immunocytochemistry staining were applied to determine the expression of NRP1 on B16F10 cells and its xenotransplanted tumors by NRP1 mAb.The proliferation,migration and apoptosis of melanoma cells were detected by MTT assay,the scratch test and Annexin V-FITC /PI double labeled flow cytometry after administration with different concentrations of NRP1 mAb.The expression level of some signal molecules was measured by Western blot.RESULTS The purified NRP1 mAb has high purity and specificity.B16F10 cells and its xenotransplanted tumors highly expressed NRP1.NRP1 mAb inhibit the proliferation and migration in a time-and dose-dependent manner( P < 0.05).Moreover NRP1 mAb induce apoptosis of B16F10 in a dose-dependent manner( P < 0.05).The results of western blot revealed down-regulated expression of p-Akt,p-ERK1 /2,Bcl-2 and caspase-3 and up-regulated Bax expression in B16F10 cells treated by NRP1 mAb.CONCLUSION The purified NRP1 mAb can effectively inhibit the proliferation and migration of B16F10 and induce apoptosis.Meanwhile,the changed expression of the proteins suggests that NRP1 mAb may inhibit cell proliferation and migration by ERK1 /2 and PI3 K /Akt pathway and induce cell apoptosis by Bcl-2 /Bax /caspase-3 pathway,consequencely playing anti-tumor role.; 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)
语种	zh_CN
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/123712]
专题	医学院－已发表论文
推荐引用方式 GB/T 7714	吕莎,李哲,王显江,等. 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, Effects of Anti-NRP1 Monoclonal Antibody on Cell Proliferation,Migration and Apoptosis of Melanoma B16F10 Cells[J],2014.
APA	吕莎.,李哲.,王显江.,王生育.,杨芸.,...&颜江华.(2014).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..
MLA	吕莎,et al."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".(2014).

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