Hsa_circ_0066523 promotes the proliferation and osteogenic differentiation of bone mesenchymal stem cells by repressing PTEN
Aims: Circular RNAs (circRNAs) are a novel type of non-coding RNA that plays major roles in the development of diverse diseases including osteonecrosis of the femoral head (ONFH). Here, we explored the impact of hsa_circ_0066523 derived from forkhead box P1 (FOXP1) (also called circFOXP1) on bone me...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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The British Editorial Society of Bone & Joint Surgery
2021-08-01
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| Series: | Bone & Joint Research |
| Subjects: | |
| Online Access: | https://online.boneandjoint.org.uk/doi/epdf/10.1302/2046-3758.108.BJR-2020-0127.R2 |
| _version_ | 1818359135477956608 |
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| author | Wei Xin Shuai Yuan Bo Wang Qirong Qian Yi Chen |
| author_facet | Wei Xin Shuai Yuan Bo Wang Qirong Qian Yi Chen |
| author_sort | Wei Xin |
| collection | DOAJ |
| description | Aims: Circular RNAs (circRNAs) are a novel type of non-coding RNA that plays major roles in the development of diverse diseases including osteonecrosis of the femoral head (ONFH). Here, we explored the impact of hsa_circ_0066523 derived from forkhead box P1 (FOXP1) (also called circFOXP1) on bone mesenchymal stem cells (BMSCs), which is important for ONFH development. Methods: RNA or protein expression in BMSCs was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) or western blot, respectively. Cell Counting Kit 8 (CCK8) and 5-ethynyl-2’-deoxyuridine (EdU) were used to analyze cell proliferation. Alkaline phosphatase (ALP) activity, ALP staining, and Alizarin Red S staining were employed to evaluate the osteoblastic differentiation. Chromatin immunoprecipitation (ChIP), luciferase reporter, RNA pull down, and RNA immunoprecipitation (RIP) assays were combined for exploring molecular associations. Results: Circ_0066523 was upregulated in osteogenic induction process of BMSCs. Silencing circ_0066523 restrained the proliferation and osteogenic differentiation of BMSCs. Mechanistically, circ_0066523 activated phosphatidylinositol-4,5-bisphosphate 3-kinase / AKT serine/threonine kinase 1 (PI3K/AKT) pathway via recruiting lysine demethylase 5B (KDM5B) to epigenetically repress the transcription of phosphatase and tensin homolog (PTEN). Functionally, AKT signalling pathway agonist or PTEN knockdown counteracted the effects of silenced circ_0066523 on BMSC proliferation and differentiation. Conclusion: Circ_0066523 promotes the proliferation and differentiation of BMSCs by epigenetically repressing PTEN and therefore activating AKT pathway. This finding might open new avenues for the identification of therapeutic targets for osteoblast differentiation related diseases such as ONFH. |
| first_indexed | 2024-12-13T20:40:05Z |
| format | Article |
| id | doaj.art-57f6b23fc7f04012aed9dddcc8a429b2 |
| institution | Directory Open Access Journal |
| issn | 2046-3758 |
| language | English |
| last_indexed | 2024-12-13T20:40:05Z |
| publishDate | 2021-08-01 |
| publisher | The British Editorial Society of Bone & Joint Surgery |
| record_format | Article |
| series | Bone & Joint Research |
| spelling | doaj.art-57f6b23fc7f04012aed9dddcc8a429b22022-12-21T23:32:11ZengThe British Editorial Society of Bone & Joint SurgeryBone & Joint Research2046-37582021-08-0110852653510.1302/2046-3758.108.BJR-2020-0127.R2Hsa_circ_0066523 promotes the proliferation and osteogenic differentiation of bone mesenchymal stem cells by repressing PTENWei Xin0Shuai Yuan1Bo Wang2Qirong Qian3Yi Chen4Department of Orthopedics, Changzheng Hospital, Shanghai, ChinaDepartment of Orthopedics, Changzheng Hospital, Shanghai, ChinaDepartment of Orthopedics, Changzheng Hospital, Shanghai, ChinaDepartment of Orthopedics, Changzheng Hospital, Shanghai, ChinaDepartment of Orthopedics, Changzheng Hospital, Shanghai, ChinaAims: Circular RNAs (circRNAs) are a novel type of non-coding RNA that plays major roles in the development of diverse diseases including osteonecrosis of the femoral head (ONFH). Here, we explored the impact of hsa_circ_0066523 derived from forkhead box P1 (FOXP1) (also called circFOXP1) on bone mesenchymal stem cells (BMSCs), which is important for ONFH development. Methods: RNA or protein expression in BMSCs was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) or western blot, respectively. Cell Counting Kit 8 (CCK8) and 5-ethynyl-2’-deoxyuridine (EdU) were used to analyze cell proliferation. Alkaline phosphatase (ALP) activity, ALP staining, and Alizarin Red S staining were employed to evaluate the osteoblastic differentiation. Chromatin immunoprecipitation (ChIP), luciferase reporter, RNA pull down, and RNA immunoprecipitation (RIP) assays were combined for exploring molecular associations. Results: Circ_0066523 was upregulated in osteogenic induction process of BMSCs. Silencing circ_0066523 restrained the proliferation and osteogenic differentiation of BMSCs. Mechanistically, circ_0066523 activated phosphatidylinositol-4,5-bisphosphate 3-kinase / AKT serine/threonine kinase 1 (PI3K/AKT) pathway via recruiting lysine demethylase 5B (KDM5B) to epigenetically repress the transcription of phosphatase and tensin homolog (PTEN). Functionally, AKT signalling pathway agonist or PTEN knockdown counteracted the effects of silenced circ_0066523 on BMSC proliferation and differentiation. Conclusion: Circ_0066523 promotes the proliferation and differentiation of BMSCs by epigenetically repressing PTEN and therefore activating AKT pathway. This finding might open new avenues for the identification of therapeutic targets for osteoblast differentiation related diseases such as ONFH.https://online.boneandjoint.org.uk/doi/epdf/10.1302/2046-3758.108.BJR-2020-0127.R2circ_0066523osteoblast differentiationkdm5bmesenchymal stem cells (mscs)bmscsrnasstainingred sosteoblastwestern blotosteonecrosis of the femoral head (onfh)quantitative real-time polymerase chain reactionphosphatase |
| spellingShingle | Wei Xin Shuai Yuan Bo Wang Qirong Qian Yi Chen Hsa_circ_0066523 promotes the proliferation and osteogenic differentiation of bone mesenchymal stem cells by repressing PTEN Bone & Joint Research circ_0066523 osteoblast differentiation kdm5b mesenchymal stem cells (mscs) bmscs rnas staining red s osteoblast western blot osteonecrosis of the femoral head (onfh) quantitative real-time polymerase chain reaction phosphatase |
| title | Hsa_circ_0066523 promotes the proliferation and osteogenic differentiation of bone mesenchymal stem cells by repressing PTEN |
| title_full | Hsa_circ_0066523 promotes the proliferation and osteogenic differentiation of bone mesenchymal stem cells by repressing PTEN |
| title_fullStr | Hsa_circ_0066523 promotes the proliferation and osteogenic differentiation of bone mesenchymal stem cells by repressing PTEN |
| title_full_unstemmed | Hsa_circ_0066523 promotes the proliferation and osteogenic differentiation of bone mesenchymal stem cells by repressing PTEN |
| title_short | Hsa_circ_0066523 promotes the proliferation and osteogenic differentiation of bone mesenchymal stem cells by repressing PTEN |
| title_sort | hsa circ 0066523 promotes the proliferation and osteogenic differentiation of bone mesenchymal stem cells by repressing pten |
| topic | circ_0066523 osteoblast differentiation kdm5b mesenchymal stem cells (mscs) bmscs rnas staining red s osteoblast western blot osteonecrosis of the femoral head (onfh) quantitative real-time polymerase chain reaction phosphatase |
| url | https://online.boneandjoint.org.uk/doi/epdf/10.1302/2046-3758.108.BJR-2020-0127.R2 |
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