Overexpression of fucosyltransferase 8 reverses the inhibitory effect of high-dose dexamethasone on osteogenic response of MC3T3-E1 preosteoblasts
Background Core fucosylation catalyzed by FUT8 is essential for TGF-β binding to TGF-β receptors. Methods Indirect TGF-β1 binding assay was used to evaluate the ability of TGF-β1 to bind to TGFBRs, Alizarin red and alkaline phosphatase staining were used to detect osteogenic differentiation and mine...
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PeerJ Inc.
2021-12-01
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| author | Zhiming Wu Tianye Lin Pan Kang Zhikun Zhuang Haibin Wang Wei He Qiushi Wei Ziqi Li |
| author_facet | Zhiming Wu Tianye Lin Pan Kang Zhikun Zhuang Haibin Wang Wei He Qiushi Wei Ziqi Li |
| author_sort | Zhiming Wu |
| collection | DOAJ |
| description | Background Core fucosylation catalyzed by FUT8 is essential for TGF-β binding to TGF-β receptors. Methods Indirect TGF-β1 binding assay was used to evaluate the ability of TGF-β1 to bind to TGFBRs, Alizarin red and alkaline phosphatase staining were used to detect osteogenic differentiation and mineralization ability , western blot and quantitative RT-PCR were used to measure the differential expression of osteogenesis-related proteins and genes. Plasmid-mediated gain-of-function study. The scale of core fucosylation modification was detected by Lectin-blot and LCA laser confocal. Results Our results showed that compared with vehicle treatment, high-dose (10−6 and 10−5 M) dexamethasone significantly inhibited cell proliferation, osteogenic differentiation, and FUT8 mRNA expression while promoting mRNA expression of adipogenesis-related genes in MC3T3-E1 cells, suggesting that downregulation of FUT8 is involved in the inhibitory effect of high-dose dexamethasone on osteogenesis. Overexpression of FUT8 significantly promoted osteogenic differentiation and activated TGF-β/Smad signaling in MC3T3-E1 cells in the presence of high-dose dexamethasone, suggesting that FUT8 reverses the inhibitory effect of high-dose dexamethasone on osteogenesis. In addition, lectin fluorescent staining and blotting showed that overexpression of FUT8 significantly reversed the inhibitory effects of high-dose dexamethasone on core fucosylation of TGFBR1 and TGFBR2. Furthermore, indirect TGF-β1 binding assay showed that overexpression of FUT8 remarkably promoted TGF-β1 binding to TGFBRs in MC3T3-E1 cells in the presence of high-dose dexamethasone. Conclusions Taken together, these results suggest that overexpression of FUT8 facilitates counteracting the inhibitory effect of dexamethasone on TGF-β signaling and osteogenesis. |
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| last_indexed | 2024-03-09T06:26:01Z |
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| spelling | doaj.art-990fd983cf2a44d5a680546c093b87a52023-12-03T11:21:13ZengPeerJ Inc.PeerJ2167-83592021-12-019e1238010.7717/peerj.12380Overexpression of fucosyltransferase 8 reverses the inhibitory effect of high-dose dexamethasone on osteogenic response of MC3T3-E1 preosteoblastsZhiming Wu0Tianye Lin1Pan Kang2Zhikun Zhuang3Haibin Wang4Wei He5Qiushi Wei6Ziqi Li7Guangzhou University of Chinese Medicine, Guangzhou, ChinaGuangzhou University of Chinese Medicine, Guangzhou, ChinaGuangzhou University of Chinese Medicine, Guangzhou, ChinaDepartment of Joint Orthopaedic, Quanzhou Orthopedic-Traumatological Hospital of Fujian Traditional Chinese Medicine University, Quanzhou, ChinaGuangzhou University of Chinese Medicine, Guangzhou, ChinaGuangzhou University of Chinese Medicine, Guangzhou, ChinaGuangzhou University of Chinese Medicine, Guangzhou, ChinaGuangzhou University of Chinese Medicine, Guangzhou, ChinaBackground Core fucosylation catalyzed by FUT8 is essential for TGF-β binding to TGF-β receptors. Methods Indirect TGF-β1 binding assay was used to evaluate the ability of TGF-β1 to bind to TGFBRs, Alizarin red and alkaline phosphatase staining were used to detect osteogenic differentiation and mineralization ability , western blot and quantitative RT-PCR were used to measure the differential expression of osteogenesis-related proteins and genes. Plasmid-mediated gain-of-function study. The scale of core fucosylation modification was detected by Lectin-blot and LCA laser confocal. Results Our results showed that compared with vehicle treatment, high-dose (10−6 and 10−5 M) dexamethasone significantly inhibited cell proliferation, osteogenic differentiation, and FUT8 mRNA expression while promoting mRNA expression of adipogenesis-related genes in MC3T3-E1 cells, suggesting that downregulation of FUT8 is involved in the inhibitory effect of high-dose dexamethasone on osteogenesis. Overexpression of FUT8 significantly promoted osteogenic differentiation and activated TGF-β/Smad signaling in MC3T3-E1 cells in the presence of high-dose dexamethasone, suggesting that FUT8 reverses the inhibitory effect of high-dose dexamethasone on osteogenesis. In addition, lectin fluorescent staining and blotting showed that overexpression of FUT8 significantly reversed the inhibitory effects of high-dose dexamethasone on core fucosylation of TGFBR1 and TGFBR2. Furthermore, indirect TGF-β1 binding assay showed that overexpression of FUT8 remarkably promoted TGF-β1 binding to TGFBRs in MC3T3-E1 cells in the presence of high-dose dexamethasone. Conclusions Taken together, these results suggest that overexpression of FUT8 facilitates counteracting the inhibitory effect of dexamethasone on TGF-β signaling and osteogenesis.https://peerj.com/articles/12380.pdfDexamethasoneOsteogenesisFucosyltransferase 8Transforming growth factor beta |
| spellingShingle | Zhiming Wu Tianye Lin Pan Kang Zhikun Zhuang Haibin Wang Wei He Qiushi Wei Ziqi Li Overexpression of fucosyltransferase 8 reverses the inhibitory effect of high-dose dexamethasone on osteogenic response of MC3T3-E1 preosteoblasts PeerJ Dexamethasone Osteogenesis Fucosyltransferase 8 Transforming growth factor beta |
| title | Overexpression of fucosyltransferase 8 reverses the inhibitory effect of high-dose dexamethasone on osteogenic response of MC3T3-E1 preosteoblasts |
| title_full | Overexpression of fucosyltransferase 8 reverses the inhibitory effect of high-dose dexamethasone on osteogenic response of MC3T3-E1 preosteoblasts |
| title_fullStr | Overexpression of fucosyltransferase 8 reverses the inhibitory effect of high-dose dexamethasone on osteogenic response of MC3T3-E1 preosteoblasts |
| title_full_unstemmed | Overexpression of fucosyltransferase 8 reverses the inhibitory effect of high-dose dexamethasone on osteogenic response of MC3T3-E1 preosteoblasts |
| title_short | Overexpression of fucosyltransferase 8 reverses the inhibitory effect of high-dose dexamethasone on osteogenic response of MC3T3-E1 preosteoblasts |
| title_sort | overexpression of fucosyltransferase 8 reverses the inhibitory effect of high dose dexamethasone on osteogenic response of mc3t3 e1 preosteoblasts |
| topic | Dexamethasone Osteogenesis Fucosyltransferase 8 Transforming growth factor beta |
| url | https://peerj.com/articles/12380.pdf |
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