期刊
JOURNAL OF PROTEOME RESEARCH
卷 20, 期 5, 页码 2195-2205出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jproteome.0c00629
关键词
proteomics; microPOTS; lithocholic acid; Barrett's esophagus; X-ray
资金
- International Research Agenda's Program of the Foundation for Polish Science [MAB/2017/03]
- European Union under the European Regional Development Fund
- Genome Canada
- Genome British Columbia
- Office of Biological and Environmental Research [grid.436923.9]
- European Regional Development Fund Project ENOCH [CZ.02.1.01/0.0/0.0/16_019/0000868]
- Ministry of Health Development of Research Organization, MH CZ - DRO (MMCI) [00209805]
The researchers utilized microPOTS to identify proteomic changes in Barrett's esophageal cells and successfully quantify differentially expressed proteins from limited numbers of cells.
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 similar to 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.
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