含神经纤毛蛋白-1靶向序列的细胞穿透肽的合成及与非小细胞肺癌的.docVIP

含神经纤毛蛋白-1靶向序列的细胞穿透肽的合成及与非小细胞肺癌的特异性结合研究 周惠君，董萍，蔡华伟，吴小艾，张文杰，庞富文，李林△ 四川大学华西医院 核医学科（成都 610041） 【摘要】目的 设计一类具有C末端基序的新型肿瘤穿透肽YCCS，并检测其对非小细胞肺癌的靶向结合能力。方法 设计了融合肺癌靶向肽段CS及神经纤毛蛋白-1靶向肽段CRMP-1的新型多肽YCCS，采用Fmoc固相合成法合成多肽，并在多肽上标记荧光素FITC，以神经纤毛蛋白-1阳性的非小细胞肺癌A549、人乳腺癌MDA-MB-231，以及神经纤毛蛋白-1阴性的正常人肺成纤维细胞HBE135-E6E7、正常人肝细胞HL-7702为研究对象，观察荧光标记的多肽与A549细胞的特异性结合能力。结果 成功设计并合成了肿瘤穿透肽FITC-YCCS用于细胞结合研究，荧光细胞结合实验证实，在5 μmol/L浓度下，YCCS多肽能特异性的与非小细胞肺癌细胞A549结合，而基本不与乳腺癌细胞及正常细胞结合。随多肽浓度升高，在20 μmol/L浓度下，YCCS多肽与乳腺癌细胞MDA-MB-231和肝细胞HL-7702出现少量结合。结论 本研究设计的肿瘤穿透肽YCCS在5 μmol/L浓度下，可检测到对非小细胞肺癌细胞A549具有特异性结合能力。 【关键词】非小细胞肺癌 肿瘤靶向 神经纤毛蛋白-1 C末端规则序列多肽 Cellular Uptake and Localization of Novel NSCLC Penetrating Peptide with Neuropilin-1 Binding Motif ZHOU Hui-jun, DONG Ping, CAI Hua-wei, WU Xiao-ai, ZHANG Wen-jie, PANG Fu-wen, LI Lin△.Department of Nuclear Medicine, West China Hospital, Sichuan University, Chengdu 610041, China △Corresponding author, E-mail: lilinhuaxi@ 【Abstract】Objective To synthesize and study the specific binding affinity of tumor-penetrating peptide YCCS to NSCLC (Non-small cell lung carcinoma) cells in vitro. Methods YCCS peptide was designed by fusing the Neuropilin-1 (NRP-1) binding sequence and NSCLC binding peptide CS. YCCS Peptide was synthesized and fluorescent labeled with N-terminal FITC. NRP-1 positive human Non-Small Cell Lung Cancer (NSCLC) cell A549, NRP-1 positive human breast cancer cell MDA-MB-231, normal human bronchial epithelium HBE135-E6E7 and human liver cell HL-7702 were incubated with 1, 5, 10 and 20 μmol/L of peptide at 37°C, respectively. After 4h incubation, the binding affinity and cellular localization of FITC-YCCS peptide were assessed by fluorescent microscope. Results After treated with 5 μmol/L peptide, significant fluorescent signals of FITC-YCCS peptide were investigated in only NSCLC A549 cells but marginal captured signal in MDA-MB-231, normal human HBE135-E6E7 or HL-7702 cells, which revealed specific NSCLC cell binding affinity. In 20 μmol/L treated group, non-specific