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Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library
Published on: April 6, 2012
LncRNA XIXT promotes osteogenic differentiation of bone mesenchymal stem cells and alleviates osteoporosis
1Department of Orthopedics, The Second Affiliated Hospital of Soochow University, Suzhou, China. qirong_dong@163.com.
Objective:
To uncover the role of XIXT in influencing the osteogenesis of hBMSCs by adsorbing microRNA-30a-5p (miRNA-30a-5p) to upregulate RUNX2.
Patients And Methods:
The serum samples were collected from osteoporosis patients and normal people. hBMSCs were isolated from femoral head tissues. The serum levels of XIXT and miRNA-30a-5p were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The expression levels and activities of the osteogenic differentiation-related genes in hBMSCs after transfection of sh-XIXT, sh-RUNX2, miRNA-30a-5p mimic, and inhibitor were detected by qRT-PCR, Western blot, ALP activity assay, and alizarin red staining. The Dual-Luciferase Reporter Gene Assay was performed to confirm the binding of XIXT to miRNA-30a-5p, as well as the binding of miRNA-30a-5p to RUNX2.
Results:
LncRNA XIXT was significantly downregulated, and miRNA-30a-5p was upregulated in the serum of osteoporosis patients. The osteogenic differentiation-related genes (ALP, RUNX2) and XIXT were markedly upregulated in a time-dependent manner, while miRNA-30a-5p level gradually decreased in hBMSCs with the prolongation of osteogenesis. The knockdown of XIXT inhibited the osteogenic differentiation of hBMSCs. In hBMSCs, XIXT regulated RUNX2 expression by targeting miRNA-30a-5p. The knockdown of miRNA-30a-5p partially reversed the inhibitory effect of XIXT on the osteogenesis of hBMSCs. However, the downregulated RUNX2 reversed the promotive effect of miRNA-30a-5p on the osteogenesis of hBMSCs.
Conclusions:
LncRNA XIXT upregulated RUNX2 by absorbing miRNA-30a-5p, and thus induced hBMSCs osteogenesis to alleviate osteoporosis.
Insights
Long non-coding RNA XIXT promotes bone formation by binding microRNA-30a-5p to increase RUNX2, potentially treating osteoporosis.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Osteoporosis is characterized by reduced bone formation.
- Long non-coding RNAs (lncRNAs) play crucial roles in cellular processes.
- The specific role of lncRNA XIXT in osteogenesis is not fully understood.
Purpose of the Study:
- To investigate the function of lncRNA XIXT in human bone marrow stem cell (hBMSC) osteogenesis.
- To elucidate the mechanism by which XIXT influences osteogenic differentiation.
- To determine the interaction between XIXT, microRNA-30a-5p, and RUNX2.
Main Methods:
- Serum samples from osteoporosis patients and healthy individuals were analyzed.
- Human bone marrow stem cells (hBMSCs) were isolated and cultured.
- Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) and Western blot were used to assess gene and protein expression.
- Alkaline phosphatase (ALP) activity and alizarin red staining were performed to evaluate osteogenic differentiation.
- Dual-luciferase reporter assays confirmed molecular interactions.
Main Results:
- LncRNA XIXT was downregulated, and microRNA-30a-5p was upregulated in osteoporosis patients' serum.
- XIXT promoted osteogenic differentiation of hBMSCs by targeting microRNA-30a-5p.
- Knockdown of XIXT inhibited osteogenesis, while microRNA-30a-5p inhibition partially rescued this effect.
- XIXT upregulates RUNX2 expression by sponging microRNA-30a-5p.
Conclusions:
- LncRNA XIXT promotes hBMSC osteogenesis by sponging microRNA-30a-5p, leading to RUNX2 upregulation.
- This mechanism suggests lncRNA XIXT as a potential therapeutic target for osteoporosis.
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