细胞穿膜肽-抗人膀胱癌单克隆抗体-载丝裂霉素白蛋白纳米微球三联体的靶向性和穿膜活性研究

Evaluation the ability of specific binding and transducting of the triad of cell penetrating peptides-human bladder cancer monoclonal antibody conjugated with MMC-loaded albumin nanoparticle

目的:探讨细胞穿膜肽(TAT-EGFP)-抗人膀胱癌单克隆抗体(BDI-1)-载丝裂霉素白蛋白纳米微球(MMC-NS)三联体的靶向性和穿膜活性。方法:采用异型双功能连接剂SPDP,偶联TAT-EGFP、BDI-1、MMCNS形成三联体偶联物,进行EJ人膀胱癌靶向结合及穿膜试验,通过激光共聚焦、扫描电子显微镜及透射电子显微镜检测其靶向结合特异性和穿膜活性。结果:成功地应用异型双功能连接剂SPDP偶联TAT-EGFP、BDI-1、载MMC白蛋白纳米微球制成三联体。TAT-EGFP-BDI-1-MMC-NS偶联物能够成功进入细胞发挥细胞毒作用。首次发现TAT-EGFP内化MMC-NS入EJ人膀胱癌细胞内部后微绒毛消失的现象。结论:TATEGFP-BDI-1-MMC-NS三联体有可能应用于膀胱肿瘤的靶向治疗。

Objective： evaluation the ability of targeting and transmembrane activity of the triad of cell penetrating peptides-human bladder cancer monoclonal antibody conjugated with MMC-loaded albumin nanoparticle. Methods： To target and penetrate bladder cancer cell by TAT-EGFP- BDI-1 -MMC-NS triplet conjugate （heter-bifunctional cross-linking agent SPDP, cell-penetrating peptides TAT-EGFP, human bladder cancer monoclonal antibody BDI-1 and MMC-loaded albumin nanoparticle ） and to test its abihty of specific binding and transmembrane activity using diaminofluorescein, under transmission electron microscope, transmission electron microscopy. Results： We succeeded in using heterbifunctional cross-linking agent SPDP, cell-penetrating peptides TAT-EGFP,human bladder cancer monoclonal antibody and MMC-loaded albumin nanoparticle to couple and form triplet conjugate（ TAT-EGFP - BDI-1 - MMC-NS）. The triad of cell penetrating peptides-human bladder cancer monoclonal antibody conjugated with MMC-loaded albumin nanoparticle suggested good cytotoxic effect on human bladder cancer cells. It was the first time to show the dependencies cancer cell membrane surface showed with microvilli disappearance after albumin nanoparticles entering cancer cells. Conchmion： This study suggested that the triad of cell penetrating peptides-human bladder cancer monoclonal antibody conjugated with MMC-Loaded Albumin Nanoparticle have potential feasibility in targeting therapy for bladder cancer.