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

Abstract

UNLABELLED:

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.

• Authors: Sheng-Ta Tsai, Chih-Chiang Tsou, Wan-Yu Mao, Wei-Chao Chang, Hsin-Ying Han, Wen-Lian Hsu, Chung-Leung Li, Chia-Ning Shen, Chung-Hsuan Chen
• Journal: Proteome science (Proteome Sci) Vol. 10 Issue 1 Pg. 69 (Nov 21 2012) ISSN: 1477-5956 [Electronic] England
• PMID: 23170877 (Publication Type: Journal Article)