表皮生长因子受体和CD13双靶点抗体药物偶联物抑制血管生成和肿瘤细胞的迁移侵袭  

Bispecific Antibody-Drug Conjugate Targeting EGFR and CD13 Inhibited Angiogenesis and Tumor Cell Migration and Invasion

作　　者：盛唯瑾[1] 弓建华[1] 江敏[1] SHENG Wei-jin;GONG Jian-hua;JIANG Min(Institute of Medicinal Biotechnology,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China)

机构地区：[1]中国医学科学院医药生物技术研究所,北京100050

出　　处：《中国药学杂志》2018年第17期1457-1462,共6页Chinese Pharmaceutical Journal

基　　金：中国医学科学院医学与健康科技创新工程资助(2016-I2M-1-011)

摘　　要：目的研究表皮生长因子受体(EGFR)/CD13双靶点抗体药物偶联物Fv-LDM-NGR抑制肿瘤生长的作用机制。方法选用人乳腺癌细胞MCF-7、人肺腺癌细胞A549和人微血管内皮细胞(HMEC)-1作为研究对象。采用四甲基偶氮唑蓝(MTT)实验测定Fv-LDM-NGR对肿瘤细胞和内皮细胞增殖的影响。小管形成(tube formation)实验观察HMEC-1细胞形成的管腔结构。Transwell实验测定Fv-LDM-NGR对肿瘤细胞迁移和侵袭的影响。Western blot分析其对细胞内EGFR细胞信号通路、细胞周期信号通路以及凋亡通路的影响。利用流式细胞术、Hoechst染色和AnnexinⅤ-FITC/PI双染法观察细胞周期的变化以及细胞凋亡。结果 Fv-LDM-NGR能够显著抑制肿瘤细胞的体外增殖,其对肿瘤细胞MCF-7和A549以及人微血管内皮细胞HMEC-1的半数抑制浓度(IC50)分别为3. 27×10^(-14)、4. 72×10^(-13)和1. 17×10^(-12)mol·L^(-1)。Fv-LDM-NGR能够干扰血管内皮细胞形成管腔结构,抑制肿瘤细胞的迁移和侵袭。对细胞信号通路具有调节作用;引起细胞周期G2/M期和S期阻滞;诱导肿瘤细胞发生凋亡。结论 Fv-LDM-NGR通过干扰CD13活性,削弱肿瘤细胞的侵袭能力,阻碍血管内皮细胞形成管腔结构;通过下调EGFR的表达及磷酸化,干扰细胞信号通路,阻滞细胞周期循环和诱导细胞凋亡,抑制肿瘤细胞的增殖和迁移。OBJECTIVE To investigate the inhibitory effects of bispecific antibody-drug conjugate Fv-LDM-NGR targeting EGFR and CD13 on human tumor cells and endothelial cells,and possible mechanisms. METHODS Human breast cancer cells MCF-7, human lung adenocarcinoma cells A549 and human microvascular endothelial cells HMEC-1, were studied. MTr assay was applied to measure proliferative activity of tumor cells. The influence of Fv-LDM-NGR on tube formation of HMEC-1 was observed. Transwell assay was applied to measure migration and invasion capacity in tumor cells. Western blot was applied for analyzing intracellular signaling transduction pathways. Flow eytometry, Hochest stain and Annexin V-FITC/PI were used to detect cell cycle and apoptosis. RESULTS Fv- LDM-NGR could inhibit the proliferation of tumor cells and mierovascular endothelial cells with IC50 values of 10-14 -10-t2mol ·L-1. Fv-LDM-NGR prevented tube formation in microvaseular endothelial cells, and suppressed migration and invasion in tumor cells. Fv- LDM-NGR interfered with the intracellular signaling transduction pathways, then caused G2/M and S phase arrest and induced apoptosis. CONCLUSION Bispecific antibody-drug conjugate Fv-LDM-NGR could prevent cell invasion in tumor cells and tube formation in microvascnlar endothelial cells through blocking activity of CD13. And it could down-regulate the expression and the phosphorylation of EG- FR, interfere with cellular signal pathways, induce cell cycle arrest and cell apoptosis, and inhibit cell proliferation and migration.

关 键 词：表皮生长因子受体 CD13 力达霉素 抗体药物偶联物 抗肿瘤 

分 类 号：R965[医药卫生—药理学]