MiR-135-5p promotes osteoblast differentiation by targeting HIF1AN in MC3T3-E1 cells
Abstract Background MicroRNAs (miRNAs or miRs) serve crucial roles in the progression of osteoporosis. This study investigated the role and specific molecular mechanism of miR-135-5p in regulating osteoblast differentiation and calcification. Methods Bone morphogenetic protein 2 (BMP2) was employed...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-08-01
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| Series: | Cellular & Molecular Biology Letters |
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| Online Access: | http://link.springer.com/article/10.1186/s11658-019-0177-6 |
| _version_ | 1818406820854628352 |
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| author | Nuo Yin Longzhang Zhu Liang Ding Junjie Yuan Li Du Mingmang Pan Feng Xue Haijun Xiao |
| author_facet | Nuo Yin Longzhang Zhu Liang Ding Junjie Yuan Li Du Mingmang Pan Feng Xue Haijun Xiao |
| author_sort | Nuo Yin |
| collection | DOAJ |
| description | Abstract Background MicroRNAs (miRNAs or miRs) serve crucial roles in the progression of osteoporosis. This study investigated the role and specific molecular mechanism of miR-135-5p in regulating osteoblast differentiation and calcification. Methods Bone morphogenetic protein 2 (BMP2) was employed to interfere with the differentiation of MC3T3-E1. Then, miR-135-5p mimic or miR-135-5p inhibitor was transfected into MC3T3-E1, and quantitative RT-PCR was used to measure the expression of miR-135-5p. The expressions of runt-related transcription factor 2 (Runx2), osterix (OSX), osteopontin (OPN), and osteocalcin (OCN) were determined using western blot. Alkaline phosphatase (ALP) activity was measured using an appropriate kit assay. Calcium nodule staining was evaluated with alizarin red staining. A luciferase reporter assay was used to verify the target of miR-135-5p. Hypoxia-inducible factor 1 α inhibitor (HIF1AN) overexpression was applied to investigate its own role in the mechanism and a miR-135-5p rescue experiment was also performed. Results Overexpression of miR-135-5p promoted osteogenic differentiation and calcification, as shown by the increase in ALP activity, calcification and osteogenic marker levels, including Runx2, OSX, OPN and OCN. Knockdown of miR-135-5p yielded the opposite results. HIF1AN was confirmed as a direct target of miR-135-5p. HIF1AN overexpression inhibited osteogenic differentiation and calcification while miR-135-5p reversed these effects. Conclusions These results indicate that miR-135-5p might have a therapeutic application related to its promotion of bone formation through the targeting of HIF1AN. |
| first_indexed | 2024-12-14T09:18:01Z |
| format | Article |
| id | doaj.art-94fc4b20fe2b46a68fe6e01cb6667699 |
| institution | Directory Open Access Journal |
| issn | 1425-8153 1689-1392 |
| language | English |
| last_indexed | 2024-12-14T09:18:01Z |
| publishDate | 2019-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Cellular & Molecular Biology Letters |
| spelling | doaj.art-94fc4b20fe2b46a68fe6e01cb66676992022-12-21T23:08:22ZengBMCCellular & Molecular Biology Letters1425-81531689-13922019-08-0124111110.1186/s11658-019-0177-6MiR-135-5p promotes osteoblast differentiation by targeting HIF1AN in MC3T3-E1 cellsNuo Yin0Longzhang Zhu1Liang Ding2Junjie Yuan3Li Du4Mingmang Pan5Feng Xue6Haijun Xiao7Department of Orthopaedics, Shanghai Fengxian District Central HospitalDepartment of Orthopaedics, Shanghai Fengxian District Central HospitalDepartment of Orthopaedics, Shanghai Fengxian District Central HospitalDepartment of Orthopaedics, Shanghai Fengxian District Central HospitalDepartment of Orthopaedics, Shanghai Fengxian District Central HospitalDepartment of Orthopaedics, Shanghai Fengxian District Central HospitalDepartment of Orthopaedics, Shanghai Fengxian District Central HospitalDepartment of Orthopaedics, Shanghai Fengxian District Central HospitalAbstract Background MicroRNAs (miRNAs or miRs) serve crucial roles in the progression of osteoporosis. This study investigated the role and specific molecular mechanism of miR-135-5p in regulating osteoblast differentiation and calcification. Methods Bone morphogenetic protein 2 (BMP2) was employed to interfere with the differentiation of MC3T3-E1. Then, miR-135-5p mimic or miR-135-5p inhibitor was transfected into MC3T3-E1, and quantitative RT-PCR was used to measure the expression of miR-135-5p. The expressions of runt-related transcription factor 2 (Runx2), osterix (OSX), osteopontin (OPN), and osteocalcin (OCN) were determined using western blot. Alkaline phosphatase (ALP) activity was measured using an appropriate kit assay. Calcium nodule staining was evaluated with alizarin red staining. A luciferase reporter assay was used to verify the target of miR-135-5p. Hypoxia-inducible factor 1 α inhibitor (HIF1AN) overexpression was applied to investigate its own role in the mechanism and a miR-135-5p rescue experiment was also performed. Results Overexpression of miR-135-5p promoted osteogenic differentiation and calcification, as shown by the increase in ALP activity, calcification and osteogenic marker levels, including Runx2, OSX, OPN and OCN. Knockdown of miR-135-5p yielded the opposite results. HIF1AN was confirmed as a direct target of miR-135-5p. HIF1AN overexpression inhibited osteogenic differentiation and calcification while miR-135-5p reversed these effects. Conclusions These results indicate that miR-135-5p might have a therapeutic application related to its promotion of bone formation through the targeting of HIF1AN.http://link.springer.com/article/10.1186/s11658-019-0177-6miR-135Osteoblast differentiationCalcificationHypoxia-inducible factor 1 α inhibitor |
| spellingShingle | Nuo Yin Longzhang Zhu Liang Ding Junjie Yuan Li Du Mingmang Pan Feng Xue Haijun Xiao MiR-135-5p promotes osteoblast differentiation by targeting HIF1AN in MC3T3-E1 cells Cellular & Molecular Biology Letters miR-135 Osteoblast differentiation Calcification Hypoxia-inducible factor 1 α inhibitor |
| title | MiR-135-5p promotes osteoblast differentiation by targeting HIF1AN in MC3T3-E1 cells |
| title_full | MiR-135-5p promotes osteoblast differentiation by targeting HIF1AN in MC3T3-E1 cells |
| title_fullStr | MiR-135-5p promotes osteoblast differentiation by targeting HIF1AN in MC3T3-E1 cells |
| title_full_unstemmed | MiR-135-5p promotes osteoblast differentiation by targeting HIF1AN in MC3T3-E1 cells |
| title_short | MiR-135-5p promotes osteoblast differentiation by targeting HIF1AN in MC3T3-E1 cells |
| title_sort | mir 135 5p promotes osteoblast differentiation by targeting hif1an in mc3t3 e1 cells |
| topic | miR-135 Osteoblast differentiation Calcification Hypoxia-inducible factor 1 α inhibitor |
| url | http://link.springer.com/article/10.1186/s11658-019-0177-6 |
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