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Updated: May 25, 2025
Laser Capture Microdissection of Mouse Embryonic Cartilage and Bone for Gene Expression Analysis
Published on: December 18, 2019
[Differential miRNA profiling and target gene analysis of marrow mesenchymal stem cells in postmenopausal
Shan-Shan Yang1, Wei Zhang, Xiao-Hua Hu
1Affiliated Stomatological Hospital of Zunyi Medical University. Zunyi 563006, Guizhou Province, China.
Purpose:
To explore the differential miRNA expression profiles and predicted target genes of mandibular bone marrow mesenchymal stem cells (MBMSCs) in a postmenopausal osteoporosis (POP) mouse model using bioinformatics methods, providing new targets for diagnosis, treatment, and prevention of POP.
Methods:
POP mouse model was established by performing ovariectomy surgery, and MBMSCs were obtained using whole bone marrow adherent culture method. Microarray sequencing was performed to detect the miRNA expression profile of MBMSCs. Subsequently, miRNA identification and prediction analysis were conducted, along with the prediction of target genes. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, and protein-protein interaction (PPI) network analysis were performed on the predicted target genes. Key hub genes were identified using algorithms such as Degree, Betweenness, and Closeness. The data were analyzed by GraphPad Prism 8.0 and R language.
Results:
A total of 84 differentially expressed miRNAs were obtained using a threshold of P＜0.05, with 33 upregulated and 51 downregulated miRNAs. GO and KEGG enrichment analyses of the 84 differentially expressed miRNAs revealed their involvement in various biological processes and pathways. Among them, 130 target gene mRNAs were enriched in the "regulation of stem cell pluripotency" signaling pathway. PPI network analysis and hub gene selection were performed for the 130 predicted target gene mRNAs, resulting in the identification of 7 reliable hub genes: Ctnnβ1, Hras, Kras, Akt1, Mapk3, Smad3, and Smad2. Among these hub genes, Ctnnβ1, Akt1, Mapk3, Smad3, and Smad2 were found to be significantly associated with POP.
Conclusions:
The differentially expressed miRNAs identified in MBMSCs of POP mice may serve as potential biomarkers and play important roles in the pathogenesis of POP. This study provides new research direction and theoretical basis for the diagnosis and treatment of POP.
