title: Changes in the methylation status of the Oct3/4, Nanog, and Sox2 promoters in stem cells during regeneration of rat tracheal epithelium after injury.
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.
authors.affiliation: Department of Pathology, College of Basic Medical Sciences, China Medical University, Shenyang, 110001, China.|Department of Pathology, First Affiliated Hospital of China Medical University, Shenyang, 110001, China.|Department of Emergency, First Affiliated Hospital of China Medical University, Shenyang, 110001, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Department of Pathology, College of Basic Medical Sciences, China Medical University, Shenyang, 110001, China.|Department of Pathology, First Affiliated Hospital of China Medical University, Shenyang, 110001, China.
authors.forename: Nan
authors.initials: N
authors.lastname: Song
authors.affiliation: Department of Pathology, College of Basic Medical Sciences, China Medical University, Shenyang, 110001, China.|Department of Pathology, First Affiliated Hospital of China Medical University, Shenyang, 110001, China.|Department of Physiology, College of Life Science and Biopharmaceutics of Shenyang Pharmaceutical University, Shenyang, 110016, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Pathology, College of Basic Medical Sciences, China Medical University, Shenyang, 110001, China.|Department of Pathology, First Affiliated Hospital of China Medical University, Shenyang, 110001, China.|Department of Pathology, Shenyang Medical College, Shenyang, 110001, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Han
authors.affiliation: Department of Pathology, College of Basic Medical Sciences, China Medical University, Shenyang, 110001, China.|Department of Pathology, First Affiliated Hospital of China Medical University, Shenyang, 110001, China.
authors.forename: Zihan
authors.initials: Z
authors.lastname: Ren
authors.affiliation: Department of Ophthalmology, The 4th Affiliated Hospital, Eye Institute, China Medical University, The Key Laboratory of Lens Research, Shenyang 110005, China.
authors.forename: Jing-Xian
authors.initials: JX
authors.lastname: Xu
authors.affiliation: Department of Pathology, College of Basic Medical Sciences, China Medical University, Shenyang, 110001, China.|Department of Pathology, First Affiliated Hospital of China Medical University, Shenyang, 110001, China.
authors.forename: Yu-Chen
authors.initials: YC
authors.lastname: Han
authors.affiliation: Department of Pathology, College of Basic Medical Sciences, China Medical University, Shenyang, 110001, China.|Department of Pathology, First Affiliated Hospital of China Medical University, Shenyang, 110001, China.|IVF Michigan, Bloomfield Hills, MI, 48304, USA.
authors.forename: Fang
authors.initials: F
authors.lastname: Li
authors.affiliation: Department of Pathology, College of Basic Medical Sciences, China Medical University, Shenyang, 110001, China.|Department of Pathology, First Affiliated Hospital of China Medical University, Shenyang, 110001, China.
authors.forename: Xinshan
authors.initials: X
authors.lastname: Jia
mesh_terms: D000818:Animals; D001374:Azacitidine / Q000494:pharmacology; D019175:DNA Methylation / Q000187:drug effects; D004195:Disease Models, Animal; D015870:Gene Expression; D007150:Immunohistochemistry; D000071317:Nanog Homeobox Protein / Q000235:genetics* / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D051381:Rats; D012038:Regeneration / Q000235:genetics*; D020545:Respiratory Mucosa / Q000502:physiology; D055748:SOXB1 Transcription Factors / Q000235:genetics* / Q000378:metabolism; D013234:Stem Cells / Q000378:metabolism*; D014132:Trachea / Q000293:injuries / Q000502:physiology; D014945:Wound Healing
chemical_list: D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; D055748:SOXB1 Transcription Factors; D001374:Azacitidine
keywords: Oct3/4; methylation; silent gene; stem cell; tracheal epithelium