1. MALDI-MS-based biomarker analysis of extracellular vesicles from human lung carcinoma cells
- Author
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Chao Zhao, Wei Li, Zitong Yu, Qian Luo, Renjie Zhang, Huitao Zhang, Hui Yang, and Shi Hu
- Subjects
0303 health sciences ,biology ,Cell growth ,Chemistry ,General Chemical Engineering ,010401 analytical chemistry ,S100A10 ,General Chemistry ,medicine.disease ,01 natural sciences ,RPS27A ,0104 chemical sciences ,Ribosome assembly ,03 medical and health sciences ,Cell culture ,Cancer cell ,biology.protein ,medicine ,Cancer research ,Lung cancer ,Fetal bovine serum ,030304 developmental biology - Abstract
Extracellular vesicles (EVs) are actively secreted by mammalian cells. They are increasingly recognized as promising circulating biomarkers of disease progression. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) is currently one of the most powerful techniques for the rapid analysis of biological samples, especially for discovering biomarkers for disease diagnosis and prognosis. It is unclear what cell culture medium components and EV isolation methods are suitable for MALDI-TOF MS analysis. Using a human lung carcinoma cell line (A549), we investigated and optimized the critical experimental conditions for EVs' protein profiling by combining differential ultracentrifugation and MALDI-TOF MS. The results demonstrated that medium components and ultracentrifugation procedures to extract EVs played important roles in MS detection. Compared with EV-depleted serum and normal serum medium, conditioned medium with 2% fetal bovine serum in this study maintained cell proliferation and displayed significant protein profiling of EVs. RPS27A (ribosomal protein), which plays an essential role in mRNA translation and ribosome assembly for the differentiation of cancer cells, was detected from the EVs of lung cancer cells associated with cancer cell migration and invasion. We also found the known tumor diagnosis marker, which is S100A10_S100 calcium-binding protein A10. Therefore, MALDI-TOF MS-based EV analysis with optimized experimental protocols can contribute to future development of rapid screening techniques of protein biomarkers associated with early cancer diagnosis.
- Published
- 2021