Differential Expression Profiling of Cell Surface Antigens of CSC like Cells Derived from a Breast Cancer Cell Line Using Mass Spectrometry

Cancer Stem Cell;Cell Surface Antigen;Biotinylation;Mass spectrometry;Protein;610

융합과학기술대학원 분자의학 및 바이오제약학과 ;

University:서울대학교 대학원

关键词: Cancer Stem Cell;    Cell Surface Antigen;    Biotinylation;    Mass spectrometry;    Protein;    610;   

Others  :  http://s-space.snu.ac.kr/bitstream/10371/133318/1/000000002966.pdf

美国|英语

来源: Seoul National University Open Repository

【 摘 要 】

Cancer stem cells (CSCs) are a specific sub-population of tumor cells that are exhibiting self-renewal ability, tumorigenesis, chemotherapy resistance and metastasis. As their unique stemness properties are enabling them to escape conventional anti-cancer therapies, CSCs have become a prime interest for the development of novel therapeutic strategies for cancer treatment. However, comprehensive CSCs surface marker discovery has been limited due to their rapid progression and rare population. Recently we have obtained CSC-like cells derived from a human breast cancer cell line. The CSC-like cells exhibit not only CSC-like properties, but they maintain undifferentiated state. To discover potential CSCs specific surface markers, we carried out in-depth analyses of cell surface protein profiling of CSC-like cells compared with the Non CSC-like cells. To improve the coverage of cell surface proteins representing a ;;snapshot” of a live cell surface, we adapted the advantages of using a cell-surface biotinylation technique followed by affinity purification, SDS-PAGE protein fractionation and mass spectrometry analysis. This strategy allowed the identification of proteins unique and up-regulated in the CSC-like cells. Among the total 1525 proteins identified, we found 94 membrane proteins including known CSC surface markers (such as CD44, Integrin β-1, CD133) that are up-regulated in the CSC-like cells compared to the Non CSC-like cells. We also identified several interesting proteins that are known to be involved in tumorigenesis, chemoresistance, and metastasis which are related with CSC properties by adapting PLGEM based signal to noise ratio into Ingenuity Pathway Analysis. We have validated those up-regulated surface antigens by immunoblot assays and found that differentially expressed membrane proteins of the CSC-like cells are related in the known functions of CSCs.