Changes in the methylation status of the Oct3/4, Nanog, and Sox2 promoters in stem cells during regeneration of rat tracheal epithelium after injury
| Autor: | Jing-xian Xu, Fang Li, Nan Song, Yuchen Han, Ying Zhou, Xin Li, Zihan Ren, Xinshan Jia, Ying Han |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Homeobox protein NANOG Methyltransferase DNMT3B Gene Expression Respiratory Mucosa silent gene Andrology 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine SOX2 Animals Regeneration Medicine Promoter Regions Genetic Wound Healing tracheal epithelium business.industry SOXB1 Transcription Factors Stem Cells Nanog Homeobox Protein Methylation DNA Methylation Immunohistochemistry Rats Oct3/4 Demethylating agent Trachea stem cell Disease Models Animal 030104 developmental biology Oncology chemistry 030220 oncology & carcinogenesis embryonic structures Immunology DNA methylation Azacitidine methylation biological phenomena cell phenomena and immunity Stem cell business Octamer Transcription Factor-3 Research Paper |
| Zdroj: | Oncotarget |
| ISSN: | 1949-2553 |
| Popis: | // Ying Zhou 1, 2, 3 , Nan Song 1, 2 , Xin Li 1, 2, 4 , Ying Han 1, 2, 5 , Zihan Ren 1, 2 , Jing-xian Xu 6 , Yu-chen Han 1, 2 , Fang Li 1, 2, 7 , Xinshan Jia 1, 2 1 Department of Pathology, College of Basic Medical Sciences, China Medical University, Shenyang, 110001, China 2 Department of Pathology, First Affiliated Hospital of China Medical University, Shenyang, 110001, China 3 Department of Emergency, First Affiliated Hospital of China Medical University, Shenyang, 110001, China 4 Department of Physiology, College of Life Science and Biopharmaceutics of Shenyang Pharmaceutical University, Shenyang, 110016, China 5 Department of Pathology, Shenyang Medical College, Shenyang, 110001, China 6 Department of Ophthalmology, The 4th Affiliated Hospital, Eye Institute, China Medical University, The Key Laboratory of Lens Research, Shenyang 110005, China 7 IVF Michigan, Bloomfield Hills, MI, 48304, USA Correspondence to: Xinshan Jia, email: xinshanjia@126.com 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. |
| Databáze: | OpenAIRE |
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