Department of Hepatobiliary-Pancreatic Surgery, China-Japan Union Hospital of Jilin University, Jilin University, No. 126 Xiantai Street, Changchun, Jilin 130000, P. R. China.
State Key Laboratory of Integrated Optoelectronic, College of Electronic Science and Engineering, Jilin University, No. 2699 Qianjin Street, Changchun 130012, P. R. China.
ACS Appl Mater Interfaces. 2022 Feb 16;14(6):7717-7730. doi: 10.1021/acsami.1c24191. Epub 2022 Feb 3.
MicroRNAs (miRNAs) are a class of small, noncoding RNAs involved in nearly all genetic central dogma processes and human biological behavior, which also play a significant role in the pathological activity of tumors, such as gene transcription, protein translation, and exosome secretion. Therefore, through the navigation of certain specific miRNAs, we can trace the specific physiological processes or image some specific tissues. Designing and accurately positioning microRNA (miRNA)-sensitive fluorescent nanoprobes with benign specificity and recognition in cells or tissues are a challenging research field. To solve the difficulties, we introduce four semiconducting polymer dots (Pdots) as nanoprobes linked by specific miRNA antisense sequences for monitoring the pathological grading by the variation in miRNA expression. Based on the base pairing principle, these miRNA-sensitive Pdots could bind to specific miRNAs within the cancerous cells. As impacted by the background of different pathology gradings, the proportions of the four hepatocellular carcinoma (HCC)-specific miRNAs within the cancerous cell are different, and the pathological grading of the patient tissues can be determined by comparing the palette combinations. The short single-stranded RNA-functionalized Pdots, which have excellent microRNA sensitivity, are observed in an experimental cell model and a series of tissue specimens from HCC patients for the first time. Using the Förster (or fluorescence) resonance energy transfer (FRET) model of Pdots and Cy3dt tag to simulate in vivo miRNA detection, the superior sensitivity and specificity of these nanoprobes are verified. The interference of subjective factors in traditional single/bis-dye emission intensity detection is abandoned, and multiple label staining is used to enhance sensitivity further and reduce the false-positive rate. The feasibility exhibited by this novel staining method is verified in normal hepatocellular HCC cell lines and 16 frozen ultrathin tissue sections, which are employed to quantify pathological grading-related color presentation systems for clinical doctors and pathologists' use. The intelligently designed miRNA-guided Pdots will emerge as an ideal platform with promising biological imaging.
微小 RNA（miRNAs）是一类小的非编码 RNA，参与几乎所有的遗传中心法则过程和人类的生物行为，在肿瘤的病理活动中也发挥着重要作用，如基因转录、蛋白质翻译和外泌体分泌。因此，通过特定的特定 miRNA 的导航，我们可以追踪特定的生理过程或对一些特定的组织成像。设计并精确定位在细胞或组织中具有良性特异性和识别性的 miRNA 敏感荧光纳米探针是一个具有挑战性的研究领域。为了解决这些困难，我们引入了四个半导体聚合物点（Pdots）作为纳米探针，通过特定 miRNA 反义序列的连接，用于监测 miRNA 表达变化的病理分级。基于碱基配对原理，这些 miRNA 敏感的 Pdots 可以与癌细胞内的特定 miRNA 结合。由于不同病理分级的背景影响，癌细胞内四种肝癌（HCC）特异性 miRNA 的比例不同，可以通过比较调色板组合来确定患者组织的病理分级。在实验细胞模型和一系列 HCC 患者的组织标本中，首次观察到具有优异 miRNA 敏感性的短单链 RNA 功能化 Pdots。利用 Pdots 和 Cy3dt 标签的福斯特（或荧光）共振能量转移（FRET）模型模拟体内 miRNA 检测，验证了这些纳米探针的优越灵敏度和特异性。摒弃了传统单/双染料发射强度检测中主观因素的干扰，采用多标签染色进一步提高了灵敏度，降低了假阳性率。该新型染色方法在正常肝细胞 HCC 细胞系和 16 个冷冻超薄组织切片中的可行性得到了验证，可用于定量病理分级相关颜色呈现系统，供临床医生和病理学家使用。这种智能设计的 miRNA 引导的 Pdots 将成为一种理想的具有广阔前景的生物成像平台。
