背景:鼻咽癌(NPC)是由鼻咽粘膜内膜引起的恶性上皮癌。在早期诊断NPC可以改善患者的预后并有助于降低癌症死亡率。癌细胞与正常细胞之间最显著的变更是细胞核的变异。因此,准确检测癌细胞与正常细胞之间细胞核的生化变化,对探索鼻咽癌诊断分子标志物具有巨大潜力。高灵敏的表面增强拉曼散射(SERS)可以在分子水平上反映细胞癌变过程中的生化变化。然而,快速核靶向SERS检测仍然是一个挑战。
结果:提出了一种基于电穿孔的新颖,准确的核靶向SERS检测方法。在电脉冲的帮助下,将核靶向纳米探针快速导入不同的NPC细胞(包括CNE1,CNE2,C666细胞系)和正常鼻咽上皮细胞(NP69细胞系),分别。在核定位信号肽(NLS)的作用下,进入细胞的纳米探针位于细胞核,提供高质量的核SERS信号。苏木精和伊红(H&E)染色和原位细胞SERS成像证实了本研究中开发的纳米探针的优异的核靶向性能。SERS信号的比较表明,NPC细胞与正常鼻咽细胞之间的生化成分存在细微差异。此外,采用SERS光谱结合主成分分析(PCA)和线性判别分析(LDA)对鼻咽癌细胞样本进行诊断和区分。灵敏度高,特异性,在从正常鼻咽上皮细胞中筛选NPC细胞时获得了准确性。
结论:据我们所知,这是首次采用核靶向SERS检测方法筛选鼻咽癌细胞的研究.基于电穿孔技术,纳米探针可以快速引入活细胞进行细胞内生化检测。核靶向SERS检测可以在分子水平上分析癌细胞核中的生化变化,这对于早期癌症筛查和抗癌药物的细胞毒性分析具有很大的潜力。
BACKGROUND: Nasopharyngeal carcinoma (NPC) is a malignant epithelial carcinoma arising from the nasopharyngeal mucosal lining. Diagnosis of NPC at early stage can improve the outcome of patients and facilitate reduction in cancer mortality. The most significant change between cancer cells and normal cells is the variation of
cell nucleus. Therefore, accurately detecting the biochemical changes in nucleus between cancer cells and normal cells has great potential to explore diagnostic molecular markers for NPC. Highly sensitive surface-enhanced Raman scattering (SERS) could reflect the biochemical changes in the process of
cell cancerization at the molecular level. However, rapid nuclear targeting SERS detection remains a challenge.
RESULTS: A novel and accurate nuclear-targeting SERS detection method based on electroporation was proposed. With the assistance of electric pulses, nuclear-targeting nanoprobes were rapidly introduced into different NPC cells (including CNE1, CNE2, C666 cell lines) and normal nasopharyngeal epithelial cells (NP69
cell line), respectively. Under the action of nuclear localization signaling peptides (NLS), the nanoprobes entering cells were located to the nucleus, providing high-quality nuclear SERS signals. Hematoxylin and eosin (H&E) staining and in situ
cell SERS imaging confirmed the excellent nuclear targeting performance of the nanoprobes developed in this study. The comparison of SERS signals indicated that there were subtle differences in the biochemical components between NPC cells and normal nasopharyngeal cells. Furthermore, SERS spectra combined with principal component analysis (PCA) and linear discriminant analysis (LDA) were employed to diagnose and distinguish NPC
cell samples, and high sensitivity, specificity, and accuracy were obtained in the screening of NPC cells from normal nasopharyngeal epithelial cells.
CONCLUSIONS: To the best of our knowledge, this is the first study that employing nuclear-targeting SERS testing to screen nasopharyngeal carcinoma cells. Based on the electroporation technology, nanoprobes can be rapidly introduced into living cells for intracellular biochemical detection. Nuclear-targeting SERS detection can analyze the biochemical changes in the nucleus of cancer cells at the molecular level, which has great potential for early cancer screening and cytotoxicity analysis of anticancer drugs.