文献类型：期刊文献

中文题名：一种用于肿瘤药物治疗的新型人源性穿膜肽的优化及其应用

英文题名：The Optimization of A Novel Human-derived Cell-penetrating Peptide Used for Anti-cancer Treatment

作者：李思[1];翟逸舟[1];陆玉婷[2];王富军[2,3];赵健[1]

机构：[1]华东理工大学生物反应器工程重点实验室,上海200237;[2]浙江日升昌药业有限公司,东阳322100;[3]上海中医药大学中药研究所,上海201203

年份：2018

卷号：38

期号：7

起止页码：40

中文期刊名：中国生物工程杂志

外文期刊名：China Biotechnology

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD2017_2018】；

基金：国家自然科学基金(81571795)资助项目

语种：中文

中文关键词：中期因子;肝素结合域;细胞穿膜肽;药物运输;抗肿瘤药物

外文关键词：Midkine;Heparin-binding domain;Cell-penetrating peptide;Drug delivery;Anti-tumor drug

摘要：细胞穿膜肽（cell-penetrating peptide,CPP）作为一种体内大分子药物跨膜运输载体被广泛研究和应用。中期因子Midkine（MK）是人体的一种带有肝素结合域（heparin-binding domain,HBD）的生长因子。报道了MK中HBD的部分富含碱性氨基酸的基因序列（命名为MK-S0）与绿色荧光蛋白（EGFP）基因融合表达后,能将EGFP有效地转运入胞内,且其穿膜转运效率高于经典穿膜肽Tat。将MK-S0序列进一步突变优化改造得到的midkine-mutantΔ4（MK-Δ4）,其穿膜效率比天然序列来源的MK-S0提高16倍以上,且MK-Δ4的穿膜转运作用适用于多种肿瘤细胞。穿膜机制分析研究结果显示,MK-Δ4可与细胞表面硫酸乙酰肝素结合,随之以巨胞饮形式内吞入胞。采用MTT方法检测的细胞生长抑制试验结果显示,连接有MK-Δ4的苦瓜来源的核糖体失活蛋白MAP30比单独的MAP30对HeLa肿瘤细胞的药效可提升5.8倍,大大提高了这种药物蛋白抑杀肿瘤细胞的效果。由此表明,源于MK的这种经过突变改造的MK-Δ4,可作为一种新型高效的细胞穿膜肽,将药物蛋白有效运输到细胞内发挥抗肿瘤效应。
Cell-penetrating peptides（ CPPs） have been widely used in decades for its ability to carry many macromolecular drugs across cell-membrane to exert their effects. Midkine（ MK） is a heparin-binding growth factor with a heparin-binding domain（ HBD）. The HBD in MK that is rich in basic amino acids（ MK-S0） was fused with enhanced green fluorescence protein（ EGFP） and then it was found that MK-S0 could deliver EGFP into cells,and its transportation capacity is much higher that classical CPPs（ such as Tat）. After the sequence optimization on MK-S0,MK-Δ4 whose trans-membrane ability was increased about 16-fold than MK-S0 was obtained. The trans-membrane ability of MK-Δ4 was also suitable for a variety of tumor cells. The further investigation of endocytic pathways on MK-Δ4 was shown that MK-Δ4 penetrates cell-membrane through interacting with heparin sulfate on the cell surface and then via macropinocytosis. The results of cell growth inhibition by MTT method showed that MK-Δ4 could enhance the inhibitory effect of a ribosome-inactivating protein-MAP30 about 5. 8-fold in HeLa cells which is significantly enhance the anti-tumor activity of MAP30. It was suggested that MK-Δ4 optimized from heparin-binding domain MK is a novel human-derived CPP with high efficiency,and is a new drug vector for anti-tumor therapy.