Abstract
BackgroundCD133-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.ResultsInitially, 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.ConclusionsThese findings confirmed that mass spectrometry-based label-free quantitative proteomics can be used to gain insights into liver cancer stem cells.
Highlights
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
Previous works have demonstrated that Huh7 hepatoma cells contain a CD133-positive subpopulation, which displays the characteristics of cancer stem cells [17,23,25]
In order to determine whether CD133+ Huh7 cells expressed characteristics of cancer stem cells” (CSCs), we performed sphere-forming experiments, TopFlash assay and drug resistance test
Summary
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. Numerous studies have identified a subpopulation of cells in a wide variety of tumors that possess stem cell characteristics, including the ability to self-renew and differentiate into heterogeneous tumor cells [1,2]. These cells are called “cancer stem cells” (CSCs) or “tumor-initiating cells” (TICs) [3]. Recent findings have revealed that the glycosylated CD133 antigen is a potential marker for the isolation of CSCs from a wide variety of tumor tissues, including glioblastomas, lung cancer, pancreatic adenocarcinomas, prostate cancer, colon cancer and hepatocellular carcinomas [8,10,12-
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