摘要
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 a activable cell-penetrating peptides(ACPPs) and to preliminarily investigate its cell-penetrating activity and subcellular distribution. Methods ACPPs was synthesized by the chemical synthesis method;the cell-penetrating activity of ACPPs was detected by the flow cytometry;the tumor-targeted cell-penetrating effect expressing ACPP was measured by the immunofluorescence and multiskan spectrum microplate spectrophotometer; the intracellular location of ACPP and the subcellular distribution of ACPPs-pc-Ad. egfp compound were examined by inverted fluorescence microscopy. Results ACPPs was successfully synthesized,the fluorescence value in the ACPPs-FITC group was much high than that in the bovine serum albumin-FITC group,the difference was statistically significant(F=4 656.600,P=0.000);ACPP had the tumor-targeted cell-penetrating activity,the fluorescence value in human lung cancer A549,cell human colon cancer cell SW480 and human ovarian cancer cell OVCAR3 was much high than that in human bronchial epithelial cell HBE,the difference was statistically significant(F=37 947.676,P=0.000);ACPP was located in cytoplasm and mediated the ACPPs-pc-Ad. egfp compound to be distributed in cytoplasm. Conclusion ACPPs is successfully synthesized,which exhibits the cell-penetrating activity and is distributed in cytoplasm.
出处
《现代医药卫生》
2016年第6期823-826,共4页
Journal of Modern Medicine & Health
基金
广州医科大学科学研究基金项目(2011C18)
关键词
肿瘤/治疗
亚细胞部分
大分子物质
细胞穿膜肽
靶向
Neoplasms/therapy
Subcellular fractions
Macromolecular substances
Activatable cellpenetrating peptide
Targeted