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

Dayeon Yu, Hyun Soo Shin, Go Choi, Yongchan Lee

Research output: Contribution to journalArticle

6 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)213-220
Number of pages8
JournalMolecular and Cellular Biochemistry
Volume396
Issue number1-2
DOIs
Publication statusPublished - 2014 Nov 1

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Proteomics
Stomach Neoplasms
Cells
Stem cells
Proteins
Neoplastic Stem Cells
Phosphopyruvate Hydratase
Electrophoresis
Protein Isoforms
Membrane Proteins
Messenger RNA
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Heat-Shock Proteins
Western Blotting
Cell Line
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Yu, Dayeon ; Shin, Hyun Soo ; Choi, Go ; Lee, Yongchan. / Proteomic analysis of CD44(+) and CD44(-) gastric cancer cells. In: Molecular and Cellular Biochemistry. 2014 ; Vol. 396, No. 1-2. pp. 213-220.
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Proteomic analysis of CD44(+) and CD44(-) gastric cancer cells. / Yu, Dayeon; Shin, Hyun Soo; Choi, Go; Lee, Yongchan.

In: Molecular and Cellular Biochemistry, Vol. 396, No. 1-2, 01.11.2014, p. 213-220.

Research output: Contribution to journalArticle

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