Sisi Zhou, Fanshu Shan, Yu Cao, Puzhen Huang, Haitang Yang, Songqin Liu
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引用次数: 0
摘要
miR-155 是一类与癌症转移和侵袭密切相关的癌症标志物。将原位检测与基因沉默相结合,不仅有助于分析细胞内microRNA表达的丰度和空间位置信息,还能起到协同治疗的作用。在这项工作中,我们利用 MCF-7 细胞膜制备了含有三种发夹 DNA 的 HD@CM 囊泡。发夹 DNA 可被内源性 miR-155 触发,从而打开自催化分子回路(ACHA),获得 Y 型 DNA 纳米结构。这种纳米结构不仅能以高灵敏度检测内源性 miR-155,进行原位成像,还能实现对细胞内 survivin mRNA 的基因调控。经 HD@CMs 检测,MDA-MB-231、MCF-7、Hela 和 HEK-293T 细胞中的 miR-155 水平分别为 7703、3978、1696 和 1229 拷贝/细胞。通过共聚焦成像发现,HD@CM 与不同细胞混合后产生的荧光与细胞内 miR-155 的含量成正比。此外,Y 型 DNA 结构的基因调控功能显著抑制了存活素蛋白的表达,细胞凋亡率高达 83%。我们期待未来将我们的 HD@CM 平台应用于临床癌症的精确诊断和可编程治疗。
In Situ Intracellular Autocatalytic Hairpin Assembly of the Y-Shaped DNA Nanostructure for miR-155 Sensing and Gene Silencing
miR-155 is a class of cancer markers closely related to cancer metastasis and invasion. Combining in situ detection with gene silencing not only helps to analyze the information on the abundance and spatial location of microRNA expression in the cell but also synergizes the therapy. In this work, we prepared HD@CM vesicles with three hairpin DNAs by using MCF-7 cell membranes. The hairpin DNAs can be triggered by endogenous miR-155, which opens the autocatalytic molecular circuit (ACHA) and obtains Y-shaped DNA nanostructures. This nanostructure not only detects endogenous miR-155 with high sensitivity for in situ imaging but also enables gene regulation of intracellular survivin mRNA. The levels of miR-155 in MDA-MB-231, MCF-7, Hela, and HEK-293T cells are found to be 7703, 3978, 1696, and 1229 copies/cell, respectively, as detected by HD@CMs. The fluorescence produced by HD@CM after coincubation with different cells is found to be proportional to the intracellular miR-155 content by confocal imaging. In addition, the gene regulatory function of the Y-shaped DNA structure resulted in significant inhibition of survivin protein expression and apoptosis rates of up to 83%. We look forward to the future application of our HD@CM platform for the precise diagnosis and programmable treatment of clinical cancers.
期刊介绍:
Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.