title: DNA Methylation Profiling in Chondrocyte Dedifferentiation In Vitro.
abstract: DNA methylation has emerged as a crucial regulator of chondrocyte dedifferentiation, which severely compromises the outcome of autologous chondrocyte implantation (ACI) treatment for cartilage defects. However, the full-scale DNA methylation profiling in chondrocyte dedifferentiation remains to be determined. Here, we performed a genome-wide DNA methylation profiling of dedifferentiated chondrocytes in monolayer culture and chondrocytes treated with DNA methylation inhibitor 5-azacytidine (5-AzaC). This research revealed that the general methylation level of CpG was increased while the COL-1A1 promoter methylation level was decreased during the chondrocyte dedifferentiation. 5-AzaC could reduce general methylation levels and reverse the chondrocyte dedifferentiation. Surprisingly, the DNA methylation level of COL-1A1 promoter was increased after 5-AzaC treatment. The COL-1A1 expression level was increased while that of SOX-9 was decreased during the chondrocyte dedifferentiation. 5-AzaC treatment up-regulated the SOX-9 expression while down-regulated the COL-1A1 promoter activity and gene expression. Taken together, these results suggested that differential regulation of the DNA methylation level of cartilage-specific genes might contribute to the chondrocyte dedifferentiation. Thus, the epigenetic manipulation of these genes could be a potential strategy to counteract the chondrocyte dedifferentiation accompanying in vitro propagation. J. Cell. Physiol. 232: 1708-1716, 2017. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Shenzhen Key Laboratory of Tissue Engineering, Department of Sports Medicine, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), Shenzhen, Guangdong Province, China.|School of Stomatology, Hainan Medical College, Haikou, Hainan Province, China.
authors.forename: Li
authors.initials: L
authors.lastname: Duan
authors.affiliation: Departments of Chemistry, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, China.|School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, Shenzhen, Guangdong Province, China.
authors.forename: Yujie
authors.initials: Y
authors.lastname: Liang
authors.affiliation: Division of Immunology, University Children's Hospital Zurich, Zurich, Switzerland.
authors.forename: Bin
authors.initials: B
authors.lastname: Ma
authors.affiliation: Shenzhen Key Laboratory of Tissue Engineering, Department of Sports Medicine, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), Shenzhen, Guangdong Province, China.
authors.forename: Daming
authors.initials: D
authors.lastname: Wang
authors.affiliation: Shenzhen Key Laboratory of Tissue Engineering, Department of Sports Medicine, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), Shenzhen, Guangdong Province, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.affiliation: Shenzhen Key Laboratory of Tissue Engineering, Department of Sports Medicine, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), Shenzhen, Guangdong Province, China.
authors.forename: Jianghong
authors.initials: J
authors.lastname: Huang
authors.affiliation: Shenzhen Key Laboratory of Tissue Engineering, Department of Sports Medicine, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), Shenzhen, Guangdong Province, China.
authors.forename: Jianyi
authors.initials: J
authors.lastname: Xiong
authors.affiliation: Shenzhen Key Laboratory of Tissue Engineering, Department of Sports Medicine, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), Shenzhen, Guangdong Province, China.
authors.forename: Liangquan
authors.initials: L
authors.lastname: Peng
authors.affiliation: Shenzhen Key Laboratory of Tissue Engineering, Department of Sports Medicine, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), Shenzhen, Guangdong Province, China.
authors.forename: Jielin
authors.initials: J
authors.lastname: Chen
authors.affiliation: Shenzhen Key Laboratory of Tissue Engineering, Department of Sports Medicine, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), Shenzhen, Guangdong Province, China.
authors.forename: Weimin
authors.initials: W
authors.lastname: Zhu
authors.affiliation: Shenzhen Key Laboratory of Tissue Engineering, Department of Sports Medicine, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), Shenzhen, Guangdong Province, China.
authors.forename: Daping
authors.initials: D
authors.lastname: Wang
mesh_terms: D000328:Adult; D001374:Azacitidine / Q000494:pharmacology; D054337:Cell Dedifferentiation / Q000187:drug effects / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics*; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D024042:Collagen Type I / Q000378:metabolism; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation / Q000187:drug effects / Q000235:genetics*; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling; D063990:Gene Ontology; D006801:Humans; D008297:Male; D010641:Phenotype; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D055758:SOX9 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
chemical_list: D024042:Collagen Type I; D055758:SOX9 Transcription Factor; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D001374:Azacitidine