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Proteomic analysis of CD44(+) and CD44(−) gastric cancer cells

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Abstract

CD44 is a cell surface protein and it is widely used as a cancer stem cell marker in various cancer types including gastric cancer. We conducted proteomic analysis in CD44(+) and CD44(−) gastric cancer cells to understand characteristics of CD44(+) and CD44(−) cells. In the present study, we sorted cells from the gastric cancer cell line MKN45 according to CD44 expression to separate out CD44(+) and CD44(−) cells. And we conducted RT-PCR to identify mRNA expression of cancer stem cell markers in CD44(+) and CD44(−) cells. Cancer stem cell markers showed upregulated expression in CD44(+) cells. Next, we performed two-dimensional electrophoresis analysis to determine the differential expression pattern of proteins in each group; control, CD44(+), and CD44(−) MKN45 cells. We found a total of 113 spots that varied in expression between CD44(+) and CD44(−) cells, and subjected 20 of those protein spots to MALDI-MS. We selected the three proteins (HSPA8; heat shock cognate 71 kDa protein isoform 1, ezrin, α-enolase) upregulated in CD44(+) cells than CD44(−) cells and one protein (prohibitin) showed increased expression in CD44(−) cells. We validated the protein expression levels of four selected proteins by Western blot. We suggest that our study could be a helpful background to study CD44(+) cancer stem-like cells and differences between CD44(+) and CD44(−) cells in gastric cancer.

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Acknowledgments

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (grant number NRF-2013R1A1A2009707) and also supported by a faculty research grant of Yonsei University College of Medicine (6-2008-0229,6-2009-0148).

Conflict of interest

The authors have no conflicts of interest to declare.

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Authors and Affiliations

  1. Department of Internal Medicine, Institute of Gastroenterology, Yonsei University College of Medicine, 134 Shinchon-Dong, Seodaemun-Ku, Seoul, 120-752, Korea

    Dayeon Yu, Go Choi & Yong Chan Lee

  2. Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea

    Dayeon Yu & Yong Chan Lee

  3. Division of Nephrology, Department of Internal Medicine, Ewha Womans University School of Medicine, Seoul, Korea

    Hyun-Soo Shin

  4. Ewha Womans University School of Medicine, Ewha Medical Research Center, Seoul, Korea

    Hyun-Soo Shin

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  1. Dayeon Yu
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  2. Hyun-Soo Shin
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Correspondence to Yong Chan Lee.

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Yu, D., Shin, HS., Choi, G. et al. Proteomic analysis of CD44(+) and CD44(−) gastric cancer cells. Mol Cell Biochem 396, 213–220 (2014). https://doi.org/10.1007/s11010-014-2156-6

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  • DOI: https://doi.org/10.1007/s11010-014-2156-6

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