Abstract：
Long-stranded non-coding RNAs (lncRNA) have important roles in disease as transcriptional regulators, mRNA processing regulators and protein synthesis factors. However, traditional methods for detecting lncRNA are time-consuming and labor-intensive, and the functions of lncRNA are still being explored. Here, we present a surface enhanced Raman spectroscopy (SERS) based biosensor for the detection of lncRNA associated with liver cancer (LC) as well as in situ cellular imaging. Using the dual SERS probes, quantitative detection of lncRNA (DAPK1-215) can be achieved with an ultra-low detection limit of 952 aM by the target-triggered assembly of core-satellite nanostructures. And the reliability of this assay can be further improved with the R2 value of 0.9923 by an internal standard probe that enables the signal dynamic calibration. Meanwhile, the high expression of DAPK1-215 mainly distributed in the cytoplasm was observed in LC cells compared with the normal ones using the SERS imaging method. Moreover, results of cellular function assays showed that DAPK1-215 promoted the migration and invasion of LC by significantly reducing the expression of the structural domain of death associated protein kinase. The development of this biosensor based on SERS can provide a sensitive and specific method for exploring the expression of lncRNA that would be a potential biomarker for the screening of LC.
摘要：
长链非编码RNA(lncRNA)作为转录调节因子在疾病中具有重要作用,mRNA加工调节剂和蛋白质合成因子。然而,传统的检测lncRNA的方法耗时耗力,lncRNA的功能仍在探索中。这里,我们提出了一种基于表面增强拉曼光谱（SERS）的生物传感器，用于检测与肝癌（LC）相关的lncRNA以及原位细胞成像。使用双SERS探头,lncRNA（DAPK1-215）的定量检测可以通过靶触发的核心卫星纳米结构的组装以952aM的超低检测限实现。并且该测定的可靠性可以通过能够进行信号动态校准的内标探针以0.9923的R2值进一步提高。同时,使用SERS成像方法，与正常细胞相比，在LC细胞中观察到DAPK1-215主要分布在细胞质中的高表达。此外，细胞功能检测结果显示,DAPK1-215通过显著降低死亡相关蛋白激酶结构域的表达,促进LC的迁移和侵袭。这种基于SERS的生物传感器的开发可以为探索lncRNA的表达提供灵敏而特异的方法，lncRNA将是筛选LC的潜在生物标志物。
