Development of next-generation ion conductance microscopy to unravel the phenotype and gene expression state of cancer cells

开发下一代离子电导显微镜以揭示癌细胞的表型和基因表达状态

基本信息

批准号：
21H01770
负责人：
Korchev Yuri.E.
金额：
$ 11.15万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2021
资助国家：
日本
起止时间：
2021-04-01 至 2025-03-31
项目状态：
未结题

项目摘要

1. We have established a new SICM quantitative nanoscale mapping technique to measure the stiffness of breast cancer MCF7 cells and skin cancer melanoma A375M cells without damaging the cells. In addition, we formed a Pt-functionalized carbon nanoelectrode at the tip of nanopipette to measure the ROS distribution of individual cancer cells. Now we can phenotype individual breast cancer MCF7 cells and skin cancer melanoma A375M cells with their stiffness and ROS distribution at single cell level.2. It is becoming clear that an acid pHe plays an essential role in cancer cell progression, invasiveness and resistance to therapy. Monitoring the pHe variation of cancer cells will provide critical information for understanding tumour heterogeneity during pathological processes such as epithelial-mesenchyme transition. Based on our previously studies on enzyme-based pH-sensing nanoprobes, we have developed non-enzyme pH-sensing nanoprobes for pHe measuring from individual cancer cells.3. Regarding gene expression evaluation, conventional methods disrupt cells and cause the instability in their results. It has been difficult to know the intracellular gene expression status of individual cancer cells. Gene sampling with SICM nanopipette can perform repeat sampling from the same cell at different time, which allows a dynamical gene expression evaluation. Under the help of SICM biopsy sampling methods, we have evaluated the mRNA expression in cytoplasm of individual cancer cells.

1。我们已经建立了一种新的SICM定量纳米级映射技术，以测量乳腺癌MCF7细胞和皮肤癌黑色素瘤A375M细胞的僵硬而不会损害细胞。此外，我们在纳米夹尖的尖端形成了PT官能化的碳纳米电极，以测量单个癌细胞的ROS分布。现在，我们可以表现出单个乳腺癌MCF7细胞和皮肤癌黑色素瘤A375M细胞，其刚度和ROS分布在单细胞水平上。2。很明显，酸PHE在癌细胞进展，侵入性和对治疗的抗性中起着至关重要的作用。监测癌细胞的PHE变异将提供关键信息，以了解病理过程中肿瘤异质性，例如上皮 - 间质转变。基于我们先前关于基于酶的pH含量纳米探针的研究，我们开发了非酶pH含量的纳米探针，用于从单个癌细胞中测量的PHE。3。关于基因表达评估，常规方法破坏了细胞并导致其结果不稳定。很难知道单个癌细胞的细胞内基因表达状态。使用SICM纳米夹的基因采样可以在不同时间进行同一细胞的重复采样，这允许动态基因表达评估。在SICM活检抽样方法的帮助下，我们评估了单个癌细胞细胞质中的mRNA表达。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Phycosphere pH of unicellular nano- and micro- phytoplankton cells and consequences for iron speciation.

DOI：
10.1038/s41396-022-01280-1
发表时间：
2022-10
期刊：
ISME JOURNAL
影响因子：
11
作者：
Liu, Fengjie;Gledhill, Martha;Tan, Qiao-Guo;Zhu, Kechen;Zhang, Qiong;Salaun, Pascal;Tagliabue, Alessandro;Zhang, Yanjun;Weiss, Dominik;Achterberg, Eric P.;Korchev, Yuri
通讯作者：
Korchev, Yuri