State Key Laboratory of Electroanalytical Chemistry, Research Center of Biomembranomics, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, 130022, China; University of Science and Technology of China, Hefei, Anhui, 230027, China.
State Key Laboratory of Electroanalytical Chemistry, Research Center of Biomembranomics, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, 130022, China.
Talanta. 2020 Sep 1;217:121037. doi: 10.1016/j.talanta.2020.121037. Epub 2020 Apr 17.
Nucleolin (NCL) is a multifunctional protein that mainly localizes in the nucleolus and also distributes in the nucleoplasm, cytoplasm and cell membrane. Most studies focus on its biofunctions in cell activities and diseases, however, its detailed distribution and organization pattern in situ remains obscure. Moreover, antibodies were commonly used to investigate NCL in cells. It is worth noting that antibody labeling of intracellular proteins needs detergents to permeabilize the membrane, which could disrupt the membrane structure and proteins. The emergence of aptamer AS1411 provides us a good choice to recognize the NCL without permeabilization owing to its superior cellular uptake and enhanced stability. Therefore, we applied aptamer AS1411 to super-resolution imaging to visualize the distribution of NCL at a nanometer level. Aptamer achieved a better recognition of intracellular NCL and displayed the detailed structure of NCL in different parts of cells. Significantly, cytoplasmic and membrane NCL have higher expression and larger clusters in cancer cells than that in normal cells. Our work presented a detailed organization of NCL in cells and revealed the distribution differences between cancer cells and normal cells, which promote the understanding of its functions in physiology and pathology.
核仁素（NCL）是一种多功能蛋白，主要定位于核仁，也分布于核质、细胞质和细胞膜。大多数研究都集中在其在细胞活动和疾病中的生物功能上，然而，其在原位的详细分布和组织模式仍不清楚。此外，抗体通常用于研究细胞中的 NCL。值得注意的是，抗体标记细胞内蛋白需要去污剂来通透细胞膜，这可能会破坏膜结构和蛋白。适配体 AS1411 的出现为我们提供了一个很好的选择，可以在不进行通透的情况下识别 NCL，因为它具有更好的细胞摄取和增强的稳定性。因此，我们应用适配体 AS1411 进行超分辨率成像，以纳米级分辨率可视化 NCL 的分布。适配体实现了对细胞内 NCL 的更好识别，并显示了细胞内不同部位 NCL 的详细结构。重要的是，细胞质和膜 NCL 在癌细胞中的表达更高，簇更大，而在正常细胞中则较低。我们的工作呈现了 NCL 在细胞中的详细组织，并揭示了癌细胞和正常细胞之间的分布差异，这有助于理解其在生理和病理中的功能。
