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研究siRNA在胞内的运输过程有助于阐明其在细胞质基质中的动力学性质及过程。本文报道了激光共聚焦扫描显微镜技术用于研究siRNA及其肽缀合物与胱氨酸骨架阳离子脂质体递送系统(siRNA/CLD)的胞内分布过程。发现siRNA在胞质内分布动力学与3′-单肽-siRNA缀合物/CLD递送系统的入胞途径紧密相关,其中具有较好抗肿瘤活性的3′-p As-siRNA/CLD,具有较快的细胞摄取及溶媒体逃逸过程,并能在A375细胞中潴留较长的时间。该方法高效、可定量并可视化,不仅能够测定siRNA在胞内的浓度,也能够定量分析其与细胞亚结构的共定位关系,进一步可系统研究siRNA及其缀合物在胞质内的动力学分布过程。
Studying the intracellular trafficking of siRNA helps clarify its kinetic properties and processes in the cytoplasmic matrix. This paper reports the confocal laser scanning microscopy technique used to study the intracellular distribution of siRNA and its peptide conjugates and the cystine cationic cationic liposome delivery system (siRNA / CLD). The distribution dynamics of siRNA in the cytoplasm was closely related to the entry pathway of 3’-monopeptide-siRNA conjugate / CLD delivery system, among which 3’-p As-siRNA / CLD had better anti-tumor activity, Faster cell uptake and solvent escape, and retained longer in A375 cells. The method is efficient, quantifiable and visualizable. It can not only determine the intracellular concentration of siRNA, but also quantitatively analyze its colocalization with cell sub-structures. The kinetics of siRNA and its conjugates in the cytoplasm can be further systematically studied Distribution process.