Epigenetic regulatory mechanism of ADAMTS12 expression in osteoarthritis
Abstract Background Osteoarthritis (OA) is a degenerative joint disease with lacking effective prevention targets. A disintegrin and metalloproteinase with thrombospondin motifs 12 (ADAMTS12) is a member of the ADAMTS family and is upregulated in OA pathologic tissues with no fully understood molecu...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-07-01
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| Series: | Molecular Medicine |
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| Online Access: | https://doi.org/10.1186/s10020-023-00661-2 |
| _version_ | 1827905410123694080 |
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| author | Shu Yang Xuanping Zhou Zhen Jia Mali Zhang Minghao Yuan Yizhao Zhou Jing Wang Duo Xia |
| author_facet | Shu Yang Xuanping Zhou Zhen Jia Mali Zhang Minghao Yuan Yizhao Zhou Jing Wang Duo Xia |
| author_sort | Shu Yang |
| collection | DOAJ |
| description | Abstract Background Osteoarthritis (OA) is a degenerative joint disease with lacking effective prevention targets. A disintegrin and metalloproteinase with thrombospondin motifs 12 (ADAMTS12) is a member of the ADAMTS family and is upregulated in OA pathologic tissues with no fully understood molecular mechanisms. Methods The anterior cruciate ligament transection (ACL-T) method was used to establish rat OA models, and interleukin-1 beta (IL-1β) was administered to induce rat chondrocyte inflammation. Cartilage damage was analyzed via hematoxylin-eosin, Periodic Acid-Schiff, safranin O-fast green, Osteoarthritis Research Society International score, and micro-computed tomography assays. Chondrocyte apoptosis was detected by flow cytometry and TdT dUTP nick-end labeling. Signal transducer and activator of transcription 1 (STAT1), ADAMTS12, and methyltransferase-like 3 (METTL3) levels were detected by immunohistochemistry, quantitative polymerase chain reaction (qPCR), western blot, or immunofluorescence assay. The binding ability was confirmed by chromatin immunoprecipitation-qPCR, electromobility shift assay, dual-luciferase reporter, or RNA immunoprecipitation (RIP) assay. The methylation level of STAT1 was analyzed by MeRIP-qPCR assay. STAT1 stability was investigated by actinomycin D assay. Results The STAT1 and ADAMTS12 expressions were significantly increased in the human and rat samples of cartilage injury, as well as in IL-1β-treated rat chondrocytes. STAT1 is bound to the promoter region of ADAMTS12 to activate its transcription. METTL3/ Insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) mediated N6-methyladenosine modification of STAT1 promoted STAT1 mRNA stability, resulting in increased expression. ADAMTS12 expression was reduced and the IL-1β-induced inflammatory chondrocyte injury was attenuated by silencing METTL3. Additionally, knocking down METTL3 in ACL-T-produced OA rats reduced ADAMTS12 expression in their cartilage tissues, thereby alleviating cartilage damage. Conclusion METTL3/IGF2BP2 axis increases STAT1 stability and expression to promote OA progression by up-regulating ADAMTS12 expression. |
| first_indexed | 2024-03-13T00:41:15Z |
| format | Article |
| id | doaj.art-0515d804e15e41dcb8caa58e3bb3c8a8 |
| institution | Directory Open Access Journal |
| issn | 1528-3658 |
| language | English |
| last_indexed | 2024-03-13T00:41:15Z |
| publishDate | 2023-07-01 |
| publisher | BMC |
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| series | Molecular Medicine |
| spelling | doaj.art-0515d804e15e41dcb8caa58e3bb3c8a82023-07-09T11:16:50ZengBMCMolecular Medicine1528-36582023-07-0129111410.1186/s10020-023-00661-2Epigenetic regulatory mechanism of ADAMTS12 expression in osteoarthritisShu Yang0Xuanping Zhou1Zhen Jia2Mali Zhang3Minghao Yuan4Yizhao Zhou5Jing Wang6Duo Xia7Department of Orthopedics, Hunan Provincial People’s Hospital (The First-affiliated Hospital of Hunan Normal University)Department of Orthopedics, The First-affiliated Hospital of Hunan Normal University (Hunan Provincial People’s Hospital)Department of Orthopedics, Hunan Provincial People’s Hospital (The First-affiliated Hospital of Hunan Normal University)Department of Orthopedics, Hunan Provincial People’s Hospital (The First-affiliated Hospital of Hunan Normal University)Department of Orthopedics, Hunan Provincial People’s Hospital (The First-affiliated Hospital of Hunan Normal University)Department of Orthopedics, Hunan Provincial People’s Hospital (The First-affiliated Hospital of Hunan Normal University)Department of Orthopedics, Hunan Provincial People’s Hospital (The First-affiliated Hospital of Hunan Normal University)Department of Orthopedics, Hunan Provincial People’s Hospital (The First-affiliated Hospital of Hunan Normal University)Abstract Background Osteoarthritis (OA) is a degenerative joint disease with lacking effective prevention targets. A disintegrin and metalloproteinase with thrombospondin motifs 12 (ADAMTS12) is a member of the ADAMTS family and is upregulated in OA pathologic tissues with no fully understood molecular mechanisms. Methods The anterior cruciate ligament transection (ACL-T) method was used to establish rat OA models, and interleukin-1 beta (IL-1β) was administered to induce rat chondrocyte inflammation. Cartilage damage was analyzed via hematoxylin-eosin, Periodic Acid-Schiff, safranin O-fast green, Osteoarthritis Research Society International score, and micro-computed tomography assays. Chondrocyte apoptosis was detected by flow cytometry and TdT dUTP nick-end labeling. Signal transducer and activator of transcription 1 (STAT1), ADAMTS12, and methyltransferase-like 3 (METTL3) levels were detected by immunohistochemistry, quantitative polymerase chain reaction (qPCR), western blot, or immunofluorescence assay. The binding ability was confirmed by chromatin immunoprecipitation-qPCR, electromobility shift assay, dual-luciferase reporter, or RNA immunoprecipitation (RIP) assay. The methylation level of STAT1 was analyzed by MeRIP-qPCR assay. STAT1 stability was investigated by actinomycin D assay. Results The STAT1 and ADAMTS12 expressions were significantly increased in the human and rat samples of cartilage injury, as well as in IL-1β-treated rat chondrocytes. STAT1 is bound to the promoter region of ADAMTS12 to activate its transcription. METTL3/ Insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) mediated N6-methyladenosine modification of STAT1 promoted STAT1 mRNA stability, resulting in increased expression. ADAMTS12 expression was reduced and the IL-1β-induced inflammatory chondrocyte injury was attenuated by silencing METTL3. Additionally, knocking down METTL3 in ACL-T-produced OA rats reduced ADAMTS12 expression in their cartilage tissues, thereby alleviating cartilage damage. Conclusion METTL3/IGF2BP2 axis increases STAT1 stability and expression to promote OA progression by up-regulating ADAMTS12 expression.https://doi.org/10.1186/s10020-023-00661-2OsteoarthritisInflammationADAMTS12STAT1METTL3IGF2BP2 |
| spellingShingle | Shu Yang Xuanping Zhou Zhen Jia Mali Zhang Minghao Yuan Yizhao Zhou Jing Wang Duo Xia Epigenetic regulatory mechanism of ADAMTS12 expression in osteoarthritis Molecular Medicine Osteoarthritis Inflammation ADAMTS12 STAT1 METTL3 IGF2BP2 |
| title | Epigenetic regulatory mechanism of ADAMTS12 expression in osteoarthritis |
| title_full | Epigenetic regulatory mechanism of ADAMTS12 expression in osteoarthritis |
| title_fullStr | Epigenetic regulatory mechanism of ADAMTS12 expression in osteoarthritis |
| title_full_unstemmed | Epigenetic regulatory mechanism of ADAMTS12 expression in osteoarthritis |
| title_short | Epigenetic regulatory mechanism of ADAMTS12 expression in osteoarthritis |
| title_sort | epigenetic regulatory mechanism of adamts12 expression in osteoarthritis |
| topic | Osteoarthritis Inflammation ADAMTS12 STAT1 METTL3 IGF2BP2 |
| url | https://doi.org/10.1186/s10020-023-00661-2 |
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