LINC01089 governs the miR-1287-5p/HSPA4 axis to negatively regulate osteogenic differentiation of mesenchymal stem cells
Aims: The involvement of long non-coding RNA (lncRNA) in bone marrow mesenchymal stem cell (MSC) osteogenic differentiation during osteoporosis (OP) development has attracted much attention. In this study, we aimed to disclose how LINC01089 functions in human mesenchymal stem cell (hMSC) osteogenic...
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The British Editorial Society of Bone & Joint Surgery
2024-12-01
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| Series: | Bone & Joint Research |
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| Online Access: | https://online.boneandjoint.org.uk/doi/epdf/10.1302/2046-3758.1312.BJR-2023-0272.R2 |
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| author | Hao Zou Fei Hu Xin Wu Bin Xu Guifeng Shang Dong An Dehao Qin Xiaolei Zhang Aofei Yang |
| author_facet | Hao Zou Fei Hu Xin Wu Bin Xu Guifeng Shang Dong An Dehao Qin Xiaolei Zhang Aofei Yang |
| author_sort | Hao Zou |
| collection | DOAJ |
| description | Aims: The involvement of long non-coding RNA (lncRNA) in bone marrow mesenchymal stem cell (MSC) osteogenic differentiation during osteoporosis (OP) development has attracted much attention. In this study, we aimed to disclose how LINC01089 functions in human mesenchymal stem cell (hMSC) osteogenic differentiation, and to study the mechanism by which LINC01089 regulates MSC osteogenesis. Methods: Quantitative reverse transcription polymerase chain reaction (RT-qPCR) and western blotting were performed to analyze LINC01089, miR-1287-5p, and heat shock protein family A (HSP70) member 4 (HSPA4) expression. The osteogenic differentiation of MSCs was assessed through alkaline phosphatase (ALP) activity, alizarin red S (ARS) staining, and by measuring the levels of osteogenic gene marker expressions using commercial kits and RT-qPCR analysis. Cell proliferative capacity was evaluated via the Cell Counting Kit-8 (CCK-8) assay. The binding of miR-1287-5p with LINC01089 and HSPA4 was verified by performing dual-luciferase reporter and RNA immunoprecipitation (RIP) experiments. Results: LINC01089 expression was reinforced in serum samples of OP patients, but it gradually diminished while hMSCs underwent osteogenic differentiation. LINC01089 knockdown facilitated hMSC osteogenic differentiation. This was substantiated by: the increase in ALP activity; ALP, runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), and osteopontin (OPN) messenger RNA (mRNA) levels; and level of ARS staining. Meanwhile, LINC01089 upregulation resulted in the opposite effects. LINC01089 targeted miR-1287-5p, and the LINC01089 knockdown-induced hMSC osteogenic differentiation was repressed by miR-1287-5p depletion. HSPA4 is a downstream function molecule of the LINC01089/miR-1287-5p pathway; miR-1287-5p negatively modulated HSPA4 levels and attenuated its functional effects. Conclusion: LINC01089 negatively regulated hMSC osteogenic differentiation, at least in part, via governing miR-1287-5p/HSPA4 signalling. These findings provide new insights into hMSC osteogenesis and bone metabolism. Cite this article: Bone Joint Res 2024;13(12):779–789. |
| format | Article |
| id | doaj-art-ed2d4ed2ffa04f8b82b769a8407899ad |
| institution | Kabale University |
| issn | 2046-3758 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | The British Editorial Society of Bone & Joint Surgery |
| record_format | Article |
| series | Bone & Joint Research |
| spelling | doaj-art-ed2d4ed2ffa04f8b82b769a8407899ad2025-01-28T06:54:06ZengThe British Editorial Society of Bone & Joint SurgeryBone & Joint Research2046-37582024-12-01131277978910.1302/2046-3758.1312.BJR-2023-0272.R2LINC01089 governs the miR-1287-5p/HSPA4 axis to negatively regulate osteogenic differentiation of mesenchymal stem cellsHao Zou0https://orcid.org/0009-0004-0002-5483Fei Hu1https://orcid.org/0009-0001-0708-9007Xin Wu2https://orcid.org/0009-0007-1498-5313Bin Xu3https://orcid.org/0009-0006-1508-1706Guifeng Shang4https://orcid.org/0009-0007-6229-2490Dong An5https://orcid.org/0009-0003-4148-7150Dehao Qin6https://orcid.org/0009-0005-6106-8038Xiaolei Zhang7https://orcid.org/0009-0005-9473-0091Aofei Yang8https://orcid.org/0000-0002-4937-3508Department of Orthopedics, Xiangyang Hospital of Integrated Traditional Chinese and Western Medicine, Xiangyang, ChinaDepartment of Orthopedics, Xiangyang Hospital of Integrated Traditional Chinese and Western Medicine, Xiangyang, ChinaDepartment of Orthopedics, Xiangyang Hospital of Integrated Traditional Chinese and Western Medicine, Xiangyang, ChinaDepartment of Orthopedics, Xiangyang Hospital of Integrated Traditional Chinese and Western Medicine, Xiangyang, ChinaDepartment of Orthopedics, Xiangyang Hospital of Integrated Traditional Chinese and Western Medicine, Xiangyang, ChinaDepartment of Orthopedics, Xiangyang Hospital of Integrated Traditional Chinese and Western Medicine, Xiangyang, ChinaDepartment of Orthopedics, Xiangyang Hospital of Integrated Traditional Chinese and Western Medicine, Xiangyang, ChinaDepartment of Orthopedics, Xiangyang Hospital of Integrated Traditional Chinese and Western Medicine, Xiangyang, ChinaDepartment of Orthopedics, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan, ChinaAims: The involvement of long non-coding RNA (lncRNA) in bone marrow mesenchymal stem cell (MSC) osteogenic differentiation during osteoporosis (OP) development has attracted much attention. In this study, we aimed to disclose how LINC01089 functions in human mesenchymal stem cell (hMSC) osteogenic differentiation, and to study the mechanism by which LINC01089 regulates MSC osteogenesis. Methods: Quantitative reverse transcription polymerase chain reaction (RT-qPCR) and western blotting were performed to analyze LINC01089, miR-1287-5p, and heat shock protein family A (HSP70) member 4 (HSPA4) expression. The osteogenic differentiation of MSCs was assessed through alkaline phosphatase (ALP) activity, alizarin red S (ARS) staining, and by measuring the levels of osteogenic gene marker expressions using commercial kits and RT-qPCR analysis. Cell proliferative capacity was evaluated via the Cell Counting Kit-8 (CCK-8) assay. The binding of miR-1287-5p with LINC01089 and HSPA4 was verified by performing dual-luciferase reporter and RNA immunoprecipitation (RIP) experiments. Results: LINC01089 expression was reinforced in serum samples of OP patients, but it gradually diminished while hMSCs underwent osteogenic differentiation. LINC01089 knockdown facilitated hMSC osteogenic differentiation. This was substantiated by: the increase in ALP activity; ALP, runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), and osteopontin (OPN) messenger RNA (mRNA) levels; and level of ARS staining. Meanwhile, LINC01089 upregulation resulted in the opposite effects. LINC01089 targeted miR-1287-5p, and the LINC01089 knockdown-induced hMSC osteogenic differentiation was repressed by miR-1287-5p depletion. HSPA4 is a downstream function molecule of the LINC01089/miR-1287-5p pathway; miR-1287-5p negatively modulated HSPA4 levels and attenuated its functional effects. Conclusion: LINC01089 negatively regulated hMSC osteogenic differentiation, at least in part, via governing miR-1287-5p/HSPA4 signalling. These findings provide new insights into hMSC osteogenesis and bone metabolism. Cite this article: Bone Joint Res 2024;13(12):779–789.https://online.boneandjoint.org.uk/doi/epdf/10.1302/2046-3758.1312.BJR-2023-0272.R2linc01089mir-1287-5phspa4osteogenic differentiationosteoporosismesenchymal stem cells (mscs)rnasstainingmrnasheat shock proteinreal-time polymerase chain reactionhuman mesenchymal stem cellsosteopontinalkaline phosphatase |
| spellingShingle | Hao Zou Fei Hu Xin Wu Bin Xu Guifeng Shang Dong An Dehao Qin Xiaolei Zhang Aofei Yang LINC01089 governs the miR-1287-5p/HSPA4 axis to negatively regulate osteogenic differentiation of mesenchymal stem cells Bone & Joint Research linc01089 mir-1287-5p hspa4 osteogenic differentiation osteoporosis mesenchymal stem cells (mscs) rnas staining mrnas heat shock protein real-time polymerase chain reaction human mesenchymal stem cells osteopontin alkaline phosphatase |
| title | LINC01089 governs the miR-1287-5p/HSPA4 axis to negatively regulate osteogenic differentiation of mesenchymal stem cells |
| title_full | LINC01089 governs the miR-1287-5p/HSPA4 axis to negatively regulate osteogenic differentiation of mesenchymal stem cells |
| title_fullStr | LINC01089 governs the miR-1287-5p/HSPA4 axis to negatively regulate osteogenic differentiation of mesenchymal stem cells |
| title_full_unstemmed | LINC01089 governs the miR-1287-5p/HSPA4 axis to negatively regulate osteogenic differentiation of mesenchymal stem cells |
| title_short | LINC01089 governs the miR-1287-5p/HSPA4 axis to negatively regulate osteogenic differentiation of mesenchymal stem cells |
| title_sort | linc01089 governs the mir 1287 5p hspa4 axis to negatively regulate osteogenic differentiation of mesenchymal stem cells |
| topic | linc01089 mir-1287-5p hspa4 osteogenic differentiation osteoporosis mesenchymal stem cells (mscs) rnas staining mrnas heat shock protein real-time polymerase chain reaction human mesenchymal stem cells osteopontin alkaline phosphatase |
| url | https://online.boneandjoint.org.uk/doi/epdf/10.1302/2046-3758.1312.BJR-2023-0272.R2 |
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