hypoxia-inducible factors activate CD133 promoter through ETS family transcription factors

Shunsuke Ohnishi; Osamu Maehara; Koji Nakagawa; Ayano Kameya; Kanako Otaki; Hirotoshi Fujita; Ryosuke Higashi; Kikuko Takagi; Masahiro Asaka; Naoya Sakamoto; Masanobu Kobayashi; Hiroshi Takeda

doi:10.1371/journal.pone.0066255

hypoxia-inducible factors activate CD133 promoter through ETS family transcription factors

PLoS One. 2013 Jun 20;8(6):e66255. doi: 10.1371/journal.pone.0066255. Print 2013.

Authors

Shunsuke Ohnishi¹, Osamu Maehara, Koji Nakagawa, Ayano Kameya, Kanako Otaki, Hirotoshi Fujita, Ryosuke Higashi, Kikuko Takagi, Masahiro Asaka, Naoya Sakamoto, Masanobu Kobayashi, Hiroshi Takeda

Affiliation

¹ Department of Gastroenterology and Hepatology, Hokkaido University Graduate School of Medicine, Sapporo, Japan. [email protected]

Abstract

CD133 is a cellular surface protein that has been reported to be a cancer stem cell marker, and thus it is considered to be a potential target for cancer treatment. However, the mechanism regulating CD133 expression is not yet understood. In this study, we analyzed the activity of five putative promoters (P1-P5) of CD133 in human embryonic kidney (HEK) 293 cells and colon cancer cell line WiDr, and found that the activity of promoters, particularly of P5, is elevated by overexpression of hypoxia-inducible factors (HIF-1α and HIF-2α). Deletion and mutation analysis identified one of the two E-twenty six (ETS) binding sites (EBSs) in the P5 region as being essential for its promoter activity induced by HIF-1α and HIF-2α. In addition, a chromatin imunoprecipitation assay demonstrated that HIF-1α and HIF-2α bind to the proximal P5 promoter at the EBSs. The immunoprecipitation assay showed that HIF-1α physically interacts with Elk1; however, HIF-2α did not bind to Elk1 or ETS1. Furthermore, knockdown of both HIF-1α and HIF-2α resulted in a reduction of CD133 expression in WiDr. Taken together, our results revealed that HIF-1α and HIF-2α activate CD133 promoter through ETS proteins.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

AC133 Antigen
Antigens, CD / genetics*
Antigens, CD / metabolism
Basic Helix-Loop-Helix Transcription Factors / physiology*
Binding Sites
Cell Hypoxia
Cell Line, Tumor
Gene Expression
Gene Knockdown Techniques
Glycoproteins / genetics*
Glycoproteins / metabolism
HEK293 Cells
Humans
Hypoxia-Inducible Factor 1, alpha Subunit / physiology*
Peptides / genetics*
Peptides / metabolism
Promoter Regions, Genetic
Protein Binding
Proto-Oncogene Protein c-ets-1 / physiology*
Transcriptional Activation*
Up-Regulation
ets-Domain Protein Elk-1 / physiology*

Substances

AC133 Antigen
Antigens, CD
Basic Helix-Loop-Helix Transcription Factors
ELK1 protein, human
ETS1 protein, human
Glycoproteins
HIF1A protein, human
Hypoxia-Inducible Factor 1, alpha Subunit
PROM1 protein, human
Peptides
Proto-Oncogene Protein c-ets-1
ets-Domain Protein Elk-1
endothelial PAS domain-containing protein 1

Grants and funding

This study was supported by Grant-in-Aid for Scientific Research (C) from Japan Society for the Promotion of Science (JSPS), and by the Yasuda Memorial Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.