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Molecular and Cellular Biochemistry

, Volume 396, Issue 1–2, pp 213–220 | Cite as

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

  • Dayeon Yu
  • Hyun-Soo Shin
  • Go Choi
  • Yong Chan Lee
Article

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.

Keywords

Cancer stem cells CD44 Gastric cancer 2-DE 

Notes

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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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Dayeon Yu
    • 1
    • 2
  • Hyun-Soo Shin
    • 3
    • 4
  • Go Choi
    • 1
  • Yong Chan Lee
    • 1
    • 2
  1. 1.Department of Internal Medicine, Institute of GastroenterologyYonsei University College of MedicineSeoulKorea
  2. 2.Brain Korea 21 PLUS Project for Medical ScienceYonsei University College of MedicineSeoulKorea
  3. 3.Division of Nephrology, Department of Internal MedicineEwha Womans University School of MedicineSeoulKorea
  4. 4.Ewha Womans University School of MedicineEwha Medical Research CenterSeoulKorea

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