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目的合成一种可活化细胞穿膜肽(ACPPs)并初步探索其穿膜活性及亚细胞分布。方法应用化学合成方法合成ACPPs,采用流式细胞仪检测ACPPs穿膜活性,免疫荧光法及全波长酶标仪检测表达ACPPs的肿瘤靶向性穿膜作用,用荧光显微镜观察ACPPs在细胞内的定位及ACPPs-pc-Ad.egfp复合物的亚细胞分布。结果成功合成了ACPPs,ACPPs-异硫氰酸荧光素(FITC)组较牛血清清蛋白-FITC组荧光量大,差异有统计学意义(F=4 656.600,P=0.000),ACPPs具有肿瘤靶向性穿膜活性,人肺癌细胞A549、人结肠癌细胞SW480、人卵巢癌细胞OVCAR3细胞质内荧光量较人支气管上皮细胞HBE大,差异有统计学意义(F=37 947.676,P=0.000);ACPPs定位于细胞质并介导ACPPs-pc-Ad.egfp复合物分布于细胞质。结论成功合成了ACPPs,ACPPs具有肿瘤靶向性穿膜活性并分布于细胞质。
Objective To synthesize an activatable cell penetrating peptide (ACPPs) and to explore its transmembrane activity and subcellular distribution. Methods ACPPs were synthesized by chemical synthesis method. The transmembrane activity of ACPPs was detected by flow cytometry. The targeting of ACPPs was detected by immunofluorescence and whole-wavelength microplate reader. The localization of ACPPs in cells was observed by fluorescence microscopy And subcellular distribution of ACPPs-pc-Ad.egfp complex. Results ACPPs were successfully synthesized. The fluorescence intensity of ACPPs-fluorescein isothiocyanate (FITC) group was higher than that of bovine serum albumin-FITC group (F = 4 656.600, P = 0.000) The trans - membrane activity of human lung cancer cell A549, human colon cancer cell SW480 and human ovarian cancer cell OVCAR3 were higher than that of human bronchial epithelial cells HBeAg (F = 37 947.676, P = 0.000). ACPPs localized in the cytoplasm and mediate the distribution of the ACPPs-pc-Ad.egfp complex in the cytoplasm. Conclusion ACPPs were successfully synthesized. ACPPs have the potential of tumor targeting and distribution in cytoplasm.