Probing the Endocytic Pathways of the Filamentous Bacteriophage in Live Cells Using Ratiometric pH Fluorescent Indicator
Probing the Endocytic Pathways of the Filamentous Bacteriophage in Live Cells Using Ratiometric pH Fluorescent Indicator
复制标题
使用比率 pH 荧光指示剂探测活细胞中丝状噬菌体的内吞途径
DOI：
10.1002/adhm.201400508
复制
发表时间：
2015-02
影响因子：
10
通讯作者：
Huang, Yong
中科院分区：
文献类型：
--
作者：
Liu, Zhi;Zhou, Quan;Niu, Zhongwei;Huang, Yong
病毒纳米颗粒在诊断和治疗的各种应用中吸引了广泛的研究兴趣。特别是，丝状M13噬菌体在生物医学应用中显示出巨大的潜力。然而，其进入细胞的途径仍不清楚，这极大地阻碍了其作为药物或基因载体的进一步应用。在这里，通过将两种荧光染料缀合到M13的表面上来设计比率M13 pH探针。由于强度比不受探针浓度、离子强度、温度、光漂白和光程长度的影响，因此该比率探针可用于研究M13的细胞内pH图。更重要的是，可以阐明M13的内化机制。发现这种丝状噬菌体在与细胞的相互作用和内化机制中表现出很大的细胞类型依赖性。噬菌体倾向于仅结合在上皮细胞而不是内皮细胞的细胞膜上。此外，M13噬菌体通过具有特定机制的内吞作用进入细胞：HeLa的网格蛋白介导的内吞作用和巨胞饮作用; MCF-7的囊泡转运、网格蛋白介导的内吞作用和巨胞饮作用;人真皮微血管内皮细胞（HDMEC）的小窝介导的内吞作用。这项工作为基于丝状噬菌体的癌症诊断和治疗提供了关键信息，特别是对于pH敏感的药物递送系统的设计。
Viral nanoparticles have attracted extensive research interests in diverse applications of diagnosis and therapy. In particular, filamentous M13 bacteriophages have shown great potential in biomedical applications. However, its pathways entering into cells still remain unclear, and this greatly hinders its further use as a drug or gene carrier. Here, a ratiometric M13 pH probe is designed by conjugating two fluorescent dyes onto the surface of M13. Since the intensity ratio is not influenced by probe concentration, ion strength, temperature, photobleaching, and optical path length, this ratiometric probe can be used to investigate the intracellular pH map of M13. More importantly, the internalization mechanism of M13 can be elucidated. It is found that this filamentous phage shows great cell‐type dependence in interaction with cells and internalization mechanism. The phage tends to be bounded on the cell membrane of only epithelial cells, not endothelial cells. Furthermore, the M13 phage enters into cells through endocytosis with specific mechanism: clathrin‐mediated endocytosis and macropinocytosis for HeLa; vesicular transport, clathrin‐mediated endocytosis, and macropinocytosis for MCF‐7; caveolae‐mediated endocytosis for human dermal microvascular endothelial cell (HDMEC). This work provides key notes for cancer diagnosis and therapy based on filamentous bacteriophage, especially for design of pH‐sensitive drug delivery systems.
登录
查看更多内容
影响因子：
14
作者：
Bhattarai, Shanta Raj;Yoo, So Young;Lee, Seung-Wuk;Dean, Deborah
通讯作者：
Dean, Deborah
影响因子：
64.8
作者：
Chung, Woo-Jae;Oh, Jin-Woo;Lee, Seung-Wuk
通讯作者：
Lee, Seung-Wuk
DOI：
10.1021/la201580v
发表时间：
2011-07
期刊：
Langmuir : the ACS journal of surfaces and colloids
影响因子：
--
作者：
Laying Wu;L. Andrew Lee;Z. Niu;S. Ghoshroy;Qian Wang
通讯作者：
Laying Wu;L. Andrew Lee;Z. Niu;S. Ghoshroy;Qian Wang
自组装噬菌体 M13 薄膜上的定向细胞生长
影响因子：
4.9
作者：
Rong, Jianhua;Lee, L. Andrew;Wang, Qian
通讯作者：
Wang, Qian
影响因子：
38.3
作者：
通讯作者：
--