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Updated: Oct 6, 2025
Induction and Analysis of Epithelial to Mesenchymal Transition
Published on: August 27, 2013
Mesenchymal-to-epithelial transition of osteoblasts induced by Fam20c knockout
Ya-Wei Geng1,2, Zhen Zhang1,2, Han Jin1,2
1Institute of Hard Tissue Development and Regeneration, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150001, Heilongjiang, People's Republic of China.
Background:
Fam20c is intimately related to tissue development and diseases. At present, it has been reported that Fam20c regulates the mineralization of osteoblasts, but there are few reports on other effects.
Objective:
To study the effect of Fam20c on osteoblasts by knocking out the Fam20c gene.
Methods:
Fam20c knockout osteoblasts were constructed by transfecting mouse osteoblasts with lentivirus. The proliferation, migration and mineralization of Fam20c knockout cells were detected by CCK-8, scratch test and alizarin red staining assays. The subcellular structure was observed by transmission electron microscopy. RT-PCR was used to detect the differential expression of mesenchymal-to-epithelial transition (MET)-related marker genes and core transcription factors. The differential expression of MET-related proteins was detected by immunofluorescence or Western blot. Transcriptome analysis of Fam20c knockout osteoblasts was performed, and real-time PCR was used to verify transcriptome analysis related to MET.
Results:
The proliferation ability of osteoblasts was not significantly changed after Fam20c deletion, but the migration ability and mineralization ability were significantly weakened. There were tight junctions between Fam20c knockout cells. The expression of mesenchymal cell marker genes and core transcription factors was significantly decreased, and the expression of epithelial cell marker genes was significantly increased. The expression of mesenchymal cell marker proteins was significantly decreased, and the expression of epithelial cell marker proteins was significantly increased. Multiple signalling molecules and pathways involved in MET have changed.
Conclusions:
Knockdown of Fam20c resulted in MET. Fam20c affects the transcription of key factors in osteoblast MET.
Insights
Fam20c gene knockout in osteoblasts impaired cell migration and mineralization, inducing mesenchymal-to-epithelial transition (MET). Fam20c influences key transcription factors in osteoblast MET.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Fam20c is crucial for tissue development and disease.
- Fam20c's role in osteoblast mineralization is known, but other functions are under-explored.
Purpose of the Study:
- To investigate the impact of Fam20c knockout on osteoblast function.
- To elucidate Fam20c's role in osteoblast mesenchymal-to-epithelial transition (MET).
Main Methods:
- Constructed Fam20c knockout osteoblasts using lentivirus transfection.
- Assessed cell proliferation, migration, and mineralization.
- Analyzed gene and protein expression related to MET using RT-PCR, Western blot, and immunofluorescence.
- Performed transcriptome analysis to identify affected pathways.
Main Results:
- Fam20c deletion significantly weakened osteoblast migration and mineralization.
- Observed tight junctions and altered expression of MET markers (decreased mesenchymal, increased epithelial).
- Identified significant changes in signaling molecules and pathways involved in MET.
Conclusions:
- Fam20c knockout induces mesenchymal-to-epithelial transition (MET) in osteoblasts.
- Fam20c plays a role in regulating the transcription of key factors in osteoblast MET.
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