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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MDAMB-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5μmol/L浓度下,YCCS多肽能特异性的与非小细胞肺癌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20μmol/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5μmol/L浓度下,可检测到对非小细胞肺癌A549具有特异性结合能力。
Objective To design a novel tumor-penetrating peptide YCCS with C-terminal motif and test its targeting ability to non-small cell lung cancer. Methods A novel peptide YCCS fused with target peptide CS of lung cancer and target peptide CRMP-1 of neuropilin-1 was designed. Fmoc solid-phase synthesis was used to synthesize the polypeptide and FITC was labeled with fluorescein FITC. -1 positive non-small cell lung cancer A549, human breast cancer MDAMB-231, and normal human fibronectin-1-negative human lung fibroblast HBE135-E6E7, normal human hepatocyte HL-7702 as the object of study to observe the fluorescently labeled Specific binding ability of polypeptide to A549 cells. Results The tumor-penetrating peptide FITC-YCCS was successfully designed and synthesized for cell-binding studies. Fluorescent cell binding experiments confirmed that the YCCS polypeptide specifically binds to non-small cell lung cancer A549 at a concentration of 5 μmol / L, Breast cancer cells and normal cells. With the increase of peptide concentration, YCCS polypeptide bound little to breast cancer cells MDA-MB-231 and hepatocyte HL-7702 at a concentration of 20μmol / L. Conclusion The tumor-penetrating peptide YCCS designed in this study could detect the specific binding ability to A549 of non-small cell lung cancer at a concentration of 5μmol / L.