Label-free quantitative proteomics of CD133-positive liver cancer stem cells

Autor: Wen-Lian Hsu, Wan Yu Mao, Sheng Ta Tsai, Wei Chao Chang, Chung-Hsuan Chen, Chia-Ning Shen, Chung Leung Li, Hsin Ying Han, Chih-Chiang Tsou
Rok vydání: 2012
Předmět:
Zdroj: Proteome Science
Proteome Science, Vol 10, Iss 1, p 69 (2012)
ISSN: 1477-5956
Popis: Background CD133-positive liver cancer stem cells, which are characterized by their resistance to conventional chemotherapy and their tumor initiation ability at limited dilutions, have been recognized as a critical target in liver cancer therapeutics. In the current work, we developed a label-free quantitative method to investigate the proteome of CD133-positive liver cancer stem cells for the purpose of identifying unique biomarkers that can be utilized for targeting liver cancer stem cells. Label-free quantitation was performed in combination with ID-based Elution time Alignment by Linear regression Quantitation (IDEAL-Q) and MaxQuant. Results Initially, IDEAL-Q analysis revealed that 151 proteins were differentially expressed in the CD133-positive hepatoma cells when compared with CD133-negative cells. We then analyzed these 151 differentially expressed proteins by MaxQuant software and identified 10 significantly up-regulated proteins. The results were further validated by RT-PCR, western blot, flow cytometry or immunofluorescent staining which revealed that prominin-1, annexin A1, annexin A3, transgelin, creatine kinase B, vimentin, and EpCAM were indeed highly expressed in the CD133-positive hepatoma cells. Conclusions These findings confirmed that mass spectrometry-based label-free quantitative proteomics can be used to gain insights into liver cancer stem cells.
Databáze: OpenAIRE
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