Single-cell microRNA sequencing method comparison and profiling of cell lines and circulating lung tumor cells

In a stepwise approach, we evaluated available protocols before performing an analysis on circulating tumor cells of small cell lung cancer patients. We first compared 19 protocols on single MCF7 cells spiked miRExplore. Second, we analyzed MCF7 single cell equivalents of the eight best protocols. Third, we carried out single cell small RNA sequencing using the best performing protocol on 8 cell lines and 67 circulating tumor cells from seven small cell lung cancer patients. Single cell miRNA-Seq /w 19 protocols, miRExplore spike-in, single cell equivalents, 8 cell lines and 67 circulating tumor cells

本研究采用分步实验策略，在针对小细胞肺癌（small cell lung cancer, SCLC）患者的循环肿瘤细胞（circulating tumor cells, CTCs）开展分析前，先对现有实验方案进行了系统性评估。首先，我们针对掺入miRExplore外参的单个MCF7细胞，对19种实验方案进行了对比；其次，针对筛选出的8种最优方案，我们对等价于单个MCF7细胞的样本开展了分析；第三，我们采用表现最佳的实验方案，对8种细胞系以及来自7名小细胞肺癌患者的67个循环肿瘤细胞实施了单细胞小RNA测序。
实验覆盖内容：19种方案的单细胞miRNA测序、miRExplore外参掺入、单细胞当量样本、8种细胞系及67个循环肿瘤细胞