Oncotarget

Research Papers:

Changes in the methylation status of the Oct3/4, Nanog, and Sox2 promoters in stem cells during regeneration of rat tracheal epithelium after injury

Ying Zhou, Nan Song, Xin Li, Ying Han, Zihan Ren, Jing-xian Xu, Yu-chen Han, Fang Li and Xinshan Jia _

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Oncotarget. 2017; 8:2984-2994. https://doi.org/10.18632/oncotarget.13818

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Abstract

Ying Zhou1,2,3, Nan Song1,2, Xin Li1,2,4, Ying Han1,2,5, Zihan Ren1,2, Jing-xian Xu6, Yu-chen Han1,2, Fang Li1,2,7, Xinshan Jia1,2

1Department of Pathology, College of Basic Medical Sciences, China Medical University, Shenyang, 110001, China

2Department of Pathology, First Affiliated Hospital of China Medical University, Shenyang, 110001, China

3Department of Emergency, First Affiliated Hospital of China Medical University, Shenyang, 110001, China

4Department of Physiology, College of Life Science and Biopharmaceutics of Shenyang Pharmaceutical University, Shenyang, 110016, China

5Department of Pathology, Shenyang Medical College, Shenyang, 110001, China

6Department of Ophthalmology, The 4th Affiliated Hospital, Eye Institute, China Medical University, The Key Laboratory of Lens Research, Shenyang 110005, China

7IVF Michigan, Bloomfield Hills, MI, 48304, USA

Correspondence to:

Xinshan Jia, email: [email protected]

Keywords: tracheal epithelium, stem cell, Oct3/4, methylation, silent gene

Received: February 25, 2016     Accepted: November 21, 2016     Published: December 07, 2016

ABSTRACT

We investigated the relationship between promoter methylation and tracheal stem cell activation. We developed a model of rat tracheal epithelium regeneration after 5-fluorouracil (5-FU)-induced injury. Using immunohistochemistry and Western blotting, the expression levels of the stem cell pluripotency regulator Oct3/4 and differentiation marker CK14 were measured after 5-FU treatment. The methylation status of the Oct3/4, Nanog, and Sox2 promoters was investigated using methylation-specific PCR. Additionally, the effects of 5-azacytidine (5-azaC), a demethylating agent, on Oct3/4, Nanog, and Sox2 mRNA and protein expression were evaluated. Finally, we measured the activity of the maintenance and de novo DNA methyltransferases DNMT1, DNMT3a, and DNMT3b. Our data indicate that Oct3/4, Sox2, and Nanog are transiently expressed in response to 5-FU-induced injury, and then they are gradually silenced as the cells differentiate. DNA methylation can result in silencing of gene expression, and it can determine whether tracheal stem cells are in an active or dormant state. Treatment with 5-FU reversed the methylation of the Oct3/4, Nanog, and Sox2 promoters, which corresponded to increases in Oct3/4, Nanog, and Sox2 mRNA and protein. Thus, both maintenance and de novo methyltransferases are involved in regulating tracheal stem cell dormancy and activation.


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