Yth m<sup>6</sup>A RNA-Binding Protein 1 Regulates Osteogenesis of MC3T3-E1 Cells under Hypoxia via Translational Control of Thrombospondin-1
Peri-implantitis is a major factor affecting implant prognosis, and the specific anatomy of the peri-implant area makes it more vulnerable to the local hypoxic environment caused by inflammation. N6-methyladenosine (m<sup>6</sup>A) plays a vital role in a multitude of biological processe...
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MDPI AG
2023-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/2/1741 |
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| author | Diwen Shi Xiaohan Liu Xinyun Li Tian Li Jie Liu Lin Wu |
| author_facet | Diwen Shi Xiaohan Liu Xinyun Li Tian Li Jie Liu Lin Wu |
| author_sort | Diwen Shi |
| collection | DOAJ |
| description | Peri-implantitis is a major factor affecting implant prognosis, and the specific anatomy of the peri-implant area makes it more vulnerable to the local hypoxic environment caused by inflammation. N6-methyladenosine (m<sup>6</sup>A) plays a vital role in a multitude of biological processes, and its main “reader” Yth m<sup>6</sup>A RNA-binding protein 1 (YTHDF1) is suggested to affect osteogenic differentiation. However, the mechanism underlying the effect of YTHDF1 on osteogenic differentiation under hypoxic conditions remains unclear. To address this question, we examined the expression of YTHDF1 under hypoxia and observed that hypoxia suppressed osteogenic differentiation but promoted the expression of YTHDF1. Then we knocked down YTHDF1 and found decreased levels of osteogenic-related markers, alkaline phosphatase (ALP) activity, and alizarin red staining (ARS) under normoxia or hypoxia treatment. Bioinformatics analysis identified Thrombospondin-1 (THBS1) might be a downstream factor of YTHDF1. The results revealed that YTHDF1 enhanced the stability of THBS1 mRNA, and immunofluorescence assays found co-localization with YTHDF1 and THBS1 under hypoxia. Loss of function studies showed knocking down YTHDF1 or THBS1 exacerbated the osteogenic inhibition caused by hypoxia. All data imply that hypoxia suppresses osteogenic differentiation and promotes the expression of YTHDF1, which translationally regulates THBS1 in an m<sup>6</sup>A-dependent manner, potentially counteracting hypoxia-induced osteogenic inhibition through the YTHDF1/THBS1 pathway. The results of this study reveal for the first time the molecular mechanism of the regulation of osteogenic differentiation by YTHDF1 under hypoxia and suggest that YTHDF1, together with its downstream factor THBS1, may be critical targets to counteract osteogenic inhibition under hypoxic conditions, providing promising therapeutic strategy for the hypoxia-induced bone loss in peri-implantitis. |
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| language | English |
| last_indexed | 2024-03-09T12:17:55Z |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-6d3ca0bcbd354b928f51f1e86e66004e2023-11-30T22:44:57ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-01-01242174110.3390/ijms24021741Yth m<sup>6</sup>A RNA-Binding Protein 1 Regulates Osteogenesis of MC3T3-E1 Cells under Hypoxia via Translational Control of Thrombospondin-1Diwen Shi0Xiaohan Liu1Xinyun Li2Tian Li3Jie Liu4Lin Wu5Department of Prosthodontics, School of Stomatology, China Medical University, Shenyang 110000, ChinaDepartment of Prosthodontics, School of Stomatology, China Medical University, Shenyang 110000, ChinaDepartment of Prosthodontics, School of Stomatology, China Medical University, Shenyang 110000, ChinaDepartment of Prosthodontics, School of Stomatology, China Medical University, Shenyang 110000, ChinaCentre of Science Experiment, China Medical University, Shenyang 110000, ChinaDepartment of Prosthodontics, School of Stomatology, China Medical University, Shenyang 110000, ChinaPeri-implantitis is a major factor affecting implant prognosis, and the specific anatomy of the peri-implant area makes it more vulnerable to the local hypoxic environment caused by inflammation. N6-methyladenosine (m<sup>6</sup>A) plays a vital role in a multitude of biological processes, and its main “reader” Yth m<sup>6</sup>A RNA-binding protein 1 (YTHDF1) is suggested to affect osteogenic differentiation. However, the mechanism underlying the effect of YTHDF1 on osteogenic differentiation under hypoxic conditions remains unclear. To address this question, we examined the expression of YTHDF1 under hypoxia and observed that hypoxia suppressed osteogenic differentiation but promoted the expression of YTHDF1. Then we knocked down YTHDF1 and found decreased levels of osteogenic-related markers, alkaline phosphatase (ALP) activity, and alizarin red staining (ARS) under normoxia or hypoxia treatment. Bioinformatics analysis identified Thrombospondin-1 (THBS1) might be a downstream factor of YTHDF1. The results revealed that YTHDF1 enhanced the stability of THBS1 mRNA, and immunofluorescence assays found co-localization with YTHDF1 and THBS1 under hypoxia. Loss of function studies showed knocking down YTHDF1 or THBS1 exacerbated the osteogenic inhibition caused by hypoxia. All data imply that hypoxia suppresses osteogenic differentiation and promotes the expression of YTHDF1, which translationally regulates THBS1 in an m<sup>6</sup>A-dependent manner, potentially counteracting hypoxia-induced osteogenic inhibition through the YTHDF1/THBS1 pathway. The results of this study reveal for the first time the molecular mechanism of the regulation of osteogenic differentiation by YTHDF1 under hypoxia and suggest that YTHDF1, together with its downstream factor THBS1, may be critical targets to counteract osteogenic inhibition under hypoxic conditions, providing promising therapeutic strategy for the hypoxia-induced bone loss in peri-implantitis.https://www.mdpi.com/1422-0067/24/2/1741hypoxiaosteoblast differentiationYTHDF1THBS1mRNA stabilityperi-implantitis |
| spellingShingle | Diwen Shi Xiaohan Liu Xinyun Li Tian Li Jie Liu Lin Wu Yth m<sup>6</sup>A RNA-Binding Protein 1 Regulates Osteogenesis of MC3T3-E1 Cells under Hypoxia via Translational Control of Thrombospondin-1 International Journal of Molecular Sciences hypoxia osteoblast differentiation YTHDF1 THBS1 mRNA stability peri-implantitis |
| title | Yth m<sup>6</sup>A RNA-Binding Protein 1 Regulates Osteogenesis of MC3T3-E1 Cells under Hypoxia via Translational Control of Thrombospondin-1 |
| title_full | Yth m<sup>6</sup>A RNA-Binding Protein 1 Regulates Osteogenesis of MC3T3-E1 Cells under Hypoxia via Translational Control of Thrombospondin-1 |
| title_fullStr | Yth m<sup>6</sup>A RNA-Binding Protein 1 Regulates Osteogenesis of MC3T3-E1 Cells under Hypoxia via Translational Control of Thrombospondin-1 |
| title_full_unstemmed | Yth m<sup>6</sup>A RNA-Binding Protein 1 Regulates Osteogenesis of MC3T3-E1 Cells under Hypoxia via Translational Control of Thrombospondin-1 |
| title_short | Yth m<sup>6</sup>A RNA-Binding Protein 1 Regulates Osteogenesis of MC3T3-E1 Cells under Hypoxia via Translational Control of Thrombospondin-1 |
| title_sort | yth m sup 6 sup a rna binding protein 1 regulates osteogenesis of mc3t3 e1 cells under hypoxia via translational control of thrombospondin 1 |
| topic | hypoxia osteoblast differentiation YTHDF1 THBS1 mRNA stability peri-implantitis |
| url | https://www.mdpi.com/1422-0067/24/2/1741 |
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