Abnormal expression of miR‑135b‑5p in bone tissue of patients with osteoporosis and its role and mechanism in osteoporosis progression

Chen,Biying; Yang,Wen; Zhao,Huiqing; Liu,Kaihua; Deng,Adi; Zhang,Guo; Pan,Kaixia

doi:10.3892/etm.2019.8278

Abnormal expression of miR‑135b‑5p in bone tissue of patients with osteoporosis and its role and mechanism in osteoporosis progression

Authors:
- Biying Chen
- Wen Yang
- Huiqing Zhao
- Kaihua Liu
- Adi Deng
- Guo Zhang
- Kaixia Pan
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Affiliations: Department of Orthopaedics, The Third Affiliated Hospital of Sun Yat‑sen University, Guangzhou, Guangdong 510530, P.R. China, Department of Rheumatology and Immunology, The Third Affiliated Hospital of Sun Yat‑sen University, Guangzhou, Guangdong 510530, P.R. China
Published online on: December 4, 2019 https://doi.org/10.3892/etm.2019.8278
Pages: 1042-1050
Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Osteoporosis (OP) is an age‑related bone disease occurring worldwide. Osteoporotic fracture is one of the leading causes of disability and death in elderly patients. MicroRNAs (miRNAs/miRs) are key molecular regulatory factors in bone remodeling processes. The present study investigated the expression and mechanism of miR‑135b‑5p in patients with osteoporosis. The present results suggested that miR‑135b‑5p was upregulated in bone tissue fragments of patients with osteoporosis compared with the control patients. MC3T3‑E1 cells were used to perform osteogenic differentiation induction. Reverse transcription‑quantitative PCR and western blot assay were used to detect the mRNA and protein expression levels of the osteogenic markers osteocalcin (OC), Osterix and alkaline phosphatase (ALP). A specific kit was used for detecting ALP activity. The present results indicated that the mRNA expression levels of OC, Osterix and ALP significantly increased on the 7 and 14th day after osteogenic differentiation induction compared with the control group. Protein expression levels of OC, Osterix and ALP also increased on the 7 and 14th day after induction. ALP assay showed that ALP activity was significantly increased on the 7 and 14th day after induction. In addition, the present study found that miR‑135b‑5p was downregulated in MC3T3‑E1 cells 7 and 14 days after osteogenic differentiation induction. The results of TargetScan analysis and dual luciferase reporter gene assay indicated that runt‑related transcription factor 2 (RUNX2) was a direct target gene of miR‑135b‑5p. RUNX2 was upregulated in MC3T3‑E1 cells on the 7 and 14th day after induction. Moreover, the present study found that compared with the osteogenic differentiation induction group, miR‑135b‑5p mimic significantly decreased OC, Osterix and ALP expression, and reduced ALP activity in MC3T3‑E1 cells. However, these reductions were reversed following overexpression of RUNX2. The present results showed that miR‑135b‑5p mimic significantly reduced cell viability in MC3T3‑E1 cells and induced cell apoptosis, and these effects were significantly reversed following RUNX2 overexpression. In summary, the present results suggested that miR‑135‑5p participated in the occurrence and development of osteoporosis via inhibition of osteogenic differentiation and osteoblast growth by targeting RUNX2. The present study suggested a novel potential target that may faciliate the treatment of osteoporosis, and further study is required to examine this possibility.

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