MiR‐149‐3p can improve the osteogenic differentiation of human adipose‐derived stem cells via targeting AKT1
Abstract The study aims to investigate the role of microRNA‐149‐3p (miR‐149‐3p) in regulating osteogenic differentiation of human adipose‐derived stem cells (hADSCs) by targeting v‐akt murine thymoma viral oncogene homolog 1 (AKT1). Bioinformatics websites and a dual luciferase reporter assay were u...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2021-12-01
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| Series: | Kaohsiung Journal of Medical Sciences |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/kjm2.12436 |
| _version_ | 1818425392474619904 |
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| author | Ai‐Ning Lai Rong Zhou Bin Chen Long Guo Yu‐Ya Dai Yong‐Peng Jia |
| author_facet | Ai‐Ning Lai Rong Zhou Bin Chen Long Guo Yu‐Ya Dai Yong‐Peng Jia |
| author_sort | Ai‐Ning Lai |
| collection | DOAJ |
| description | Abstract The study aims to investigate the role of microRNA‐149‐3p (miR‐149‐3p) in regulating osteogenic differentiation of human adipose‐derived stem cells (hADSCs) by targeting v‐akt murine thymoma viral oncogene homolog 1 (AKT1). Bioinformatics websites and a dual luciferase reporter assay were used to predict and verify the targeting relationship between miR‐149‐3p and AKT1. The hADSCs were divided into the blank, negative control (NC), mimic, control siRNA, AKT1 siRNA, and miR‐149‐3p inhibitors + AKT1 siRNA groups and then subjected to Alizarin Red staining, Alkaline phosphatase (ALP) staining, ALP activity detections, MTT assay, and EdU cell proliferation assay. Gene or protein expression was quantified using quantitative real‐time PCR (qRT‐PCR) or Western blotting, respectively. The miR‐149‐3p expression increased gradually and AKT1 expression decreased gradually during osteogenic differentiation of hADSCs. The prediction of bioinformatics websites miRTarBase and TargetScan and the dual luciferase reporter assay indicated that miR‐149‐3p can directly target AKT1. After hADSCs were transfected with miR‐149‐3p mimic, AKT1 expression was significantly downregulated. However, transfection with AKT1 siRNA did not have an impact on miR‐149‐3p in hADSCs. In comparison with the AKT1 siRNA group, the miR‐149‐3p inhibitors + AKT1 siRNA group showed decreased miR‐149‐3p expression but increased AKT1 expression. In addition, AKT1 siRNA enhanced the cell viability and proliferation of hADSCs and increased mineral calcium deposition and ALP activity, resulting in higher expression of osteogenic differentiation‐related genes, which was reversed by miR‐149‐3p inhibition. The miR‐149‐3p can increase the expression of osteogenic differentiation‐related genes by targeting AKT1 and thereby enhance the osteogenic differentiation of hADSCs. |
| first_indexed | 2024-12-14T14:13:13Z |
| format | Article |
| id | doaj.art-470af756e7f945f2b1ddd86a222bfd45 |
| institution | Directory Open Access Journal |
| issn | 1607-551X 2410-8650 |
| language | English |
| last_indexed | 2024-12-14T14:13:13Z |
| publishDate | 2021-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | Kaohsiung Journal of Medical Sciences |
| spelling | doaj.art-470af756e7f945f2b1ddd86a222bfd452022-12-21T22:58:15ZengWileyKaohsiung Journal of Medical Sciences1607-551X2410-86502021-12-0137121077108810.1002/kjm2.12436MiR‐149‐3p can improve the osteogenic differentiation of human adipose‐derived stem cells via targeting AKT1Ai‐Ning Lai0Rong Zhou1Bin Chen2Long Guo3Yu‐Ya Dai4Yong‐Peng Jia5Section II, Department of Orthopaedics The 72nd Army Hospital of PLA Zhejiang ChinaSection II, Department of Orthopaedics The 72nd Army Hospital of PLA Zhejiang ChinaSection II, Department of Orthopaedics The 72nd Army Hospital of PLA Zhejiang ChinaSection II, Department of Orthopaedics The 72nd Army Hospital of PLA Zhejiang ChinaSection II, Department of Orthopaedics The 72nd Army Hospital of PLA Zhejiang ChinaSection V, Department of Orthopaedics The 72nd Army Hospital of PLA Zhejiang ChinaAbstract The study aims to investigate the role of microRNA‐149‐3p (miR‐149‐3p) in regulating osteogenic differentiation of human adipose‐derived stem cells (hADSCs) by targeting v‐akt murine thymoma viral oncogene homolog 1 (AKT1). Bioinformatics websites and a dual luciferase reporter assay were used to predict and verify the targeting relationship between miR‐149‐3p and AKT1. The hADSCs were divided into the blank, negative control (NC), mimic, control siRNA, AKT1 siRNA, and miR‐149‐3p inhibitors + AKT1 siRNA groups and then subjected to Alizarin Red staining, Alkaline phosphatase (ALP) staining, ALP activity detections, MTT assay, and EdU cell proliferation assay. Gene or protein expression was quantified using quantitative real‐time PCR (qRT‐PCR) or Western blotting, respectively. The miR‐149‐3p expression increased gradually and AKT1 expression decreased gradually during osteogenic differentiation of hADSCs. The prediction of bioinformatics websites miRTarBase and TargetScan and the dual luciferase reporter assay indicated that miR‐149‐3p can directly target AKT1. After hADSCs were transfected with miR‐149‐3p mimic, AKT1 expression was significantly downregulated. However, transfection with AKT1 siRNA did not have an impact on miR‐149‐3p in hADSCs. In comparison with the AKT1 siRNA group, the miR‐149‐3p inhibitors + AKT1 siRNA group showed decreased miR‐149‐3p expression but increased AKT1 expression. In addition, AKT1 siRNA enhanced the cell viability and proliferation of hADSCs and increased mineral calcium deposition and ALP activity, resulting in higher expression of osteogenic differentiation‐related genes, which was reversed by miR‐149‐3p inhibition. The miR‐149‐3p can increase the expression of osteogenic differentiation‐related genes by targeting AKT1 and thereby enhance the osteogenic differentiation of hADSCs.https://doi.org/10.1002/kjm2.12436AKT1hADSCmiR‐149‐3posteogenic differentiation |
| spellingShingle | Ai‐Ning Lai Rong Zhou Bin Chen Long Guo Yu‐Ya Dai Yong‐Peng Jia MiR‐149‐3p can improve the osteogenic differentiation of human adipose‐derived stem cells via targeting AKT1 Kaohsiung Journal of Medical Sciences AKT1 hADSC miR‐149‐3p osteogenic differentiation |
| title | MiR‐149‐3p can improve the osteogenic differentiation of human adipose‐derived stem cells via targeting AKT1 |
| title_full | MiR‐149‐3p can improve the osteogenic differentiation of human adipose‐derived stem cells via targeting AKT1 |
| title_fullStr | MiR‐149‐3p can improve the osteogenic differentiation of human adipose‐derived stem cells via targeting AKT1 |
| title_full_unstemmed | MiR‐149‐3p can improve the osteogenic differentiation of human adipose‐derived stem cells via targeting AKT1 |
| title_short | MiR‐149‐3p can improve the osteogenic differentiation of human adipose‐derived stem cells via targeting AKT1 |
| title_sort | mir 149 3p can improve the osteogenic differentiation of human adipose derived stem cells via targeting akt1 |
| topic | AKT1 hADSC miR‐149‐3p osteogenic differentiation |
| url | https://doi.org/10.1002/kjm2.12436 |
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