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MiRNA-205 modulates cellular invasion and migration via regulating zinc finger E-box binding homeobox 2 expression in esophageal squamous cell carcinoma cells

Kayoko Matsushima1, Hajime Isomoto12*, Naoyuki Yamaguchi12, Naoki Inoue1, Haruhisa Machida1, Toshiyuki Nakayama3, Tomayoshi Hayashi3, Masaki Kunizaki4, Shigekazu Hidaka4, Takeshi Nagayasu4, Masahiro Nakashima5, Kenta Ujifuku6, Norisato Mitsutake6, Akira Ohtsuru6, Shunichi Yamashita6, Manav Korpal7, Yibin Kang7, Philip A Gregory8, Gregory J Goodall8, Shigeru Kohno2 and Kazuhiko Nakao1

Author Affiliations

1 Department of Gastroenterology and Hepatology, Nagasaki University Hospital, 1-7-1 Sakamoto, Nagasaki 852-8102, Japan

2 Department of Endoscopy, Nagasaki University Hospital, 1-7-1 Sakamoto, Nagasaki 852-8102, Japan

3 Department of Pathology, Nagasaki University Hospital, 1-7-1 Sakamoto, Nagasaki 852-8102, Japan

4 First Department of Surgery, Nagasaki University Hospital, 1-7-1 Sakamoto, Nagasaki 852-8102, Japan

5 Division of Tumor and Diagnostic Pathology, Atomic Bomb Disease Institute, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8102, Japan

6 Department of Molecular Medicine, Atomic Bomb Disease Institute, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8102, Japan

7 Department of Molecular Biology, Princeton University, Washington Road, Princeton, NJ 08544-1014 USA

8 Hanson Institute and Division of Human Immunology, Institute of Medical and Veterinary Science, Frome Rd, Adelaide, SA 5000, Australia

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Journal of Translational Medicine 2011, 9:30  doi:10.1186/1479-5876-9-30

Published: 22 March 2011

Abstract

Background

Esophageal squamous cell carcinoma (ESCC) is often diagnosed at later stages until they are incurable. MicroRNA (miR) is a small, non-coding RNA that negatively regulates gene expression mainly via translational repression. Accumulating evidence indicates that deregulation of miR is associated with human malignancies including ESCC. The aim of this study was to identify miR that could be specifically expressed and exert distinct biological actions in ESCC.

Methods

Total RNA was extracted from ESCC cell lines, OE21 and TE10, and a non-malignant human esophageal squamous cell line, Het-1A, and subjected to microarray analysis. Expression levels of miR that showed significant differences between the 2 ESCC and Het-1A cells based on the comprehensive analysis were analyzed by the quantitative reverse transcriptase (RT)-PCR method. Then, functional analyses, including cellular proliferation, apoptosis and Matrigel invasion and the wound healing assay, for the specific miR were conducted. Using ESCC tumor samples and paired surrounding non-cancerous tissue obtained endoscopically, the association with histopathological differentiation was examined with quantitative RT-PCR.

Results

Based on the miR microarray analysis, there were 14 miRs that showed significant differences (more than 2-fold) in expression between the 2 ESCC cells and non-malignant Het-1A. Among the significantly altered miRs, miR-205 expression levels were exclusively higher in 5 ESCC cell lines examined than any other types of malignant cell lines and Het-1A. Thus, miR-205 could be a specific miR in ESCC. Modulation of miR-205 expression by transfection with its precursor or anti-miR-205 inhibitor did not affect ESCC cell proliferation and apoptosis, but miR-205 was found to be involved in cell invasion and migration. Western blot revealed that knockdown of miR-205 expression in ESCC cells substantially enhanced expression of zinc finger E-box binding homeobox 2, accompanied by reduction of E-cadherin, a regulator of epithelial mesenchymal transition. The miR-205 expression levels were not associated with histological differentiation of human ESCC.

Conclusions

These results imply that miR-205 is an ESCC-specific miR that exerts tumor-suppressive activities with EMT inhibition by targeting ZEB2.