可活化细胞穿膜肽的设计及修饰高分子载体pHPMA的作用检测

目的 合成一种可活化细胞穿膜肽ACPP，并初步探索其穿膜活性及其对高分子聚合物pHPMA的共价修饰作用。方 法通过化学合成的方法合成可活化细胞穿膜肽ACPP，通过免疫荧光法检测细胞内mmp2蛋白的表达；通过荧光显微镜观察鉴定ACPP的穿膜活性。通过化学修饰法合成接合物pc-Ad.egfp 及ACPP-pc-Ad.egfp。通过荧光显微镜观察及动态光散射法初步鉴定接合物的合成，通过荧光显微镜观察鉴定接合物ACPP-pc-Ad.egfp 亚细胞分布。结果成功合成了可活化细胞穿膜肽ACPP；ACPP具有肿瘤靶向性穿膜活性；通过ACPP修饰高分子聚合物pHPMA合成了ACPP-pc-Ad.egfp；经荧光显微镜观察证实ACPP-pc-Ad.egfp 具有较强的感染细胞的能力，并由ACPP介导大分子进行非内吞性跨膜运输。结论成功合成了ACPP，ACPP 具有肿瘤靶向性穿膜活性，并成功修饰高分子聚合物pHPMA。

Study on synthesis of ACPP peptide and its effect on covalent modification of HPMApolymer

ObjectiveTo achieve a novel macromolecules to traverse cell membranes by synthesis and characterization of a tumor- targeted ACPP and an ACPP modified- HPMA polymer conjugates. Methods Activable cell- penetrating peptide (ACPP) were prepared by free radical precipitation copolymerization method, the expression of mmp2 was measured by Immunofluorescence; the tumor-targeted cell-penetrating ability of ACPP was verified by inverted fluorescence microscopy; ACPP- pc- Ad.egfp conjugates was checked by inverted fluorescence microscopy and dynamic light scattering (DLS); Intracellular distribution of ACPP-pc-Ad. egfp was examined by inverted fluorescence microscopy. ResultsPeptides ACPP was synthesized as devised and ACPP targeted and penetrated the tumor cells. inverted fluorescence microscopy and DLS assays demonstrated the formation of ACPP-pc- Ad.egfp conjugates; ACPP-pc- Ad.egfp showed strong ability to translocate into tumor cells exhibited nonendocytotic cytoplasmic delivery. Conclusiontumo- targeted ACPP was synthesized and modified the HPMApolymer successfully.