Analysis of an alternative human CD133 promoter reveals the implication of Ras/ERK pathway in tumor stem-like hallmarks

Kouichi Tabu, Taichi Kimura, Ken Sasai, Lei Wang, Norihisa Bizen, Hiroshi Nishihara, Tetsuya Taga, Shinya Tanaka

Research output: Contribution to journalArticle

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Abstract

Background: An increasing number of studies support the presence of stem-like cells in human malignancies. These cells are primarily responsible for tumor initiation and thus considered as a potential target to eradicate tumors. CD133 has been identified as an important cell surface marker to enrich the stem-like population in various human tumors. To reveal the molecular machinery underlying the stem-like features in tumor cells, we analyzed a promoter of CD133 gene using human colon carcinoma Caco-2 and synovial sarcoma Fuji cells, which endogenously express CD133 gene.Results: A reporter analysis revealed that P5 promoter, located far upstream in a human CD133 gene locus, exhibits the highest activity among the five putative promoters (P1 to P5). Deletion and mutation analysis identified two ETS binding sites in the P5 region as being essential for its promoter activity. Electrophoretic mobility shift assays demonstrated the specific binding between nuclear factors and the ETS binding sequence. Overexpression of dominant-negative forms of Ets2 and Elk1 resulted in the significant decrease of P5 activity. Furthermore, treatment of Fuji cells with a specific MEK/ERK inhibitor, U0126, also markedly decreased CD133 expression, but there was no significant effect in Caco-2 cells, suggesting cell type-specific regulation of CD133 expression. Instead, the side population, another hallmark of TSLCs, was dramatically diminished in Caco-2 cells by U0126. Finally, Ras-mediated oncogenic transformation in normal human astrocytes conferred the stem-like capability to form neurosphere-like colonies with the increase of CD133 mRNA expression.Conclusions: In conclusion, the Ras/ERK pathway at least in part contributes to the maintenance and the acquisition of stem-like hallmarks, although the extent of its contribution is varied in a cell type-specific manner. These findings could help our comprehensive understanding of tumor stemness, and also improve the development of eradicative therapies against human malignancies.

Original languageEnglish
Article number39
JournalMolecular Cancer
Volume9
DOIs
Publication statusPublished - 2010 Feb 19
Externally publishedYes

Fingerprint

MAP Kinase Signaling System
Neoplasms
Caco-2 Cells
Genes
Synovial Sarcoma
Sequence Deletion
Mitogen-Activated Protein Kinase Kinases
Electrophoretic Mobility Shift Assay
Astrocytes
Population
Colon
Stem Cells
Binding Sites
Maintenance
Carcinoma
Messenger RNA

ASJC Scopus subject areas

  • Molecular Medicine
  • Oncology
  • Cancer Research

Cite this

Analysis of an alternative human CD133 promoter reveals the implication of Ras/ERK pathway in tumor stem-like hallmarks. / Tabu, Kouichi; Kimura, Taichi; Sasai, Ken; Wang, Lei; Bizen, Norihisa; Nishihara, Hiroshi; Taga, Tetsuya; Tanaka, Shinya.

In: Molecular Cancer, Vol. 9, 39, 19.02.2010.

Research output: Contribution to journalArticle

Tabu, Kouichi ; Kimura, Taichi ; Sasai, Ken ; Wang, Lei ; Bizen, Norihisa ; Nishihara, Hiroshi ; Taga, Tetsuya ; Tanaka, Shinya. / Analysis of an alternative human CD133 promoter reveals the implication of Ras/ERK pathway in tumor stem-like hallmarks. In: Molecular Cancer. 2010 ; Vol. 9.
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AU - Bizen, Norihisa

AU - Nishihara, Hiroshi

AU - Taga, Tetsuya

AU - Tanaka, Shinya

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