Moving from macroscale preparative systems in proteomics to micro- and nanotechnologies offers researchers the ability to deeply profile smaller numbers of cells that are more likely to be encountered in clinical settings. Herein a recently developed microscale proteomic method, microdroplet processing in one pot for trace samples (microPOTS), was employed to identify proteomic changes in ∼200 Barrett’s esophageal cells following physiologic and radiation stress exposure. From this small population of cells, microPOTS confidently identified >1500 protein groups, and achieved a high reproducibility with a Pearson’s correlation coefficient value of R > 0.9 and over 50% protein overlap from replicates. A Barrett’s cell line model treated with either lithocholic acid (LCA) or X-ray had 21 (e.g., ASNS, RALY, FAM120A, UBE2M, IDH1, ESD) and 32 (e.g., GLUL, CALU, SH3BGRL3, S100A9, FKBP3, AGR2) overexpressed proteins, respectively, compared to the untreated set. These results demonstrate the ability of microPOTS to routinely identify and quantify differentially expressed proteins from limited numbers of cells.

中文翻译：

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Barrett食管细胞系模型的MicroPOTS分析可识别生理和辐射应激后的蛋白质组变化

从蛋白质组学的大规模制备系统到微米和纳米技术，研究人员能够深入剖析较少数量的细胞，而这些细胞在临床环境中更可能会遇到。在本文中，最近开发了一种微型蛋白质组学方法，即在一个罐中对微量样品（microPOTS）进行微滴处理，以鉴定生理和辐射应力暴露后约200巴雷特食管细胞中的蛋白质组学变化。microPOTS可以从这小部分细胞中可靠地鉴定出1500个以上的蛋白质组，并以R的Pearson相关系数值实现了高重现性。> 0.9，重复样本中的蛋白质重叠超过50％。用石胆酸（LCA）或X射线处理的Barrett细胞系模型具有21个（例如，ASNS，RALY，FAM120A，UBE2M，IDH1，ESD）和32个（例如，GLUL，CALU，SH3BGRL3，S100A9，FKBP3，AGR2） ）分别与未处理的蛋白质组相比过表达的蛋白质。这些结果证明了microPOTS能够从有限数量的细胞中常规鉴定和定量表达差异蛋白的能力。