Related Experiment Video
Updated: Dec 31, 2025
Impedance-based Real-time Measurement of Cancer Cell Migration and Invasion
Published on: April 2, 2020
MiR-194-5p inhibited metastasis and EMT of nephroblastoma cells through targeting Crk
Hong Liu1, Shi-Ying Ren2, Yan Qu3
1Department of Pediatric Surgery, Hospital Affiliated to Zunyi Medical University, Zunyi City, China.
Abstract:
Wilms tumor (WT) is the most common solid childhood tumors all over the world. MicroRNAs (miRs) contribute to tumorigenesis of various cancers through targeting gene. The present study investigated the vital role of miR-194-5p and its underlying mechanism in the progression of WT. Immunohistochemistry and quantitative real-time polymerase chain reaction (qRT-PCR) assay indicated downregulation of miR194-5p and upregulation of Crk, in WT tissues compared to adjacent normal tissues. Transfection with miR-194-5p mimics into nephroblastoma cells showed a significant decline in cell migration and invasion, which was detected by Transwell assay. Luciferase assay confirmed that Crk was a direct target gene of miR-194-5p. More important, the mesenchymal to epithelial transition (EMT) biomarkers containing E-cadherin, N-cadherin and Zeb1 were examined by Western blot, and revealed that miR-194-5p mimics decreased the levels of N-cadherin and Zeb1 but increased E-cadherin, which suggested that miR-194-5p inhibited EMT. Crk knockdown could reverse the increased nephroblastoma cell invasion, migration and EMT caused by miR-194-5p inhibitor. Interestingly, qRT-PCR and Western blot analysis showed that overexpression of miR-194-5p deactivated HGF/c-Met/Scr signaling pathway via targeting Crk. In conclusion, miR-194-5p inhibited nephroblastoma cell metastasis and EMT in the progression of WT by targeting Crk. Thus, miR-194-5p might be a potential target in WT particularly for the prevention of metastasis and EMT.
Insights
MicroRNA-194-5p (miR-194-5p) suppresses Wilms tumor (WT) progression by inhibiting cell migration and epithelial-mesenchymal transition (EMT). This microRNA targets Crk, impacting the HGF/c-Met/Scr pathway, suggesting miR-194-5p as a potential therapeutic target for WT metastasis.
Area of Science:
- Oncology
- Molecular Biology
- Gene Regulation
Background:
- Wilms tumor (WT) is the most common pediatric solid malignancy.
- MicroRNAs (miRs) are key regulators in cancer development, often by targeting specific genes.
- Understanding the role of miRs in WT tumorigenesis is crucial for developing novel therapeutic strategies.
Purpose of the Study:
- To investigate the role and mechanism of miR-194-5p in the progression of Wilms tumor (WT).
- To identify the direct target gene of miR-194-5p and its downstream signaling pathways involved in WT.
- To evaluate the potential of miR-194-5p as a therapeutic target for preventing WT metastasis and epithelial-mesenchymal transition (EMT).
Main Methods:
- Immunohistochemistry and quantitative real-time polymerase chain reaction (qRT-PCR) to assess miR-194-5p and Crk expression in WT tissues.
- Transwell assays to evaluate cell migration and invasion.
- Luciferase reporter assays to confirm direct targeting of Crk by miR-194-5p.
- Western blot analysis to examine EMT biomarkers (E-cadherin, N-cadherin, Zeb1) and signaling pathway components.
- Cell transfection with miR-194-5p mimics and Crk knockdown.
Main Results:
- miR-194-5p was significantly downregulated, while Crk was upregulated in WT tissues compared to normal adjacent tissues.
- Overexpression of miR-194-5p reduced nephroblastoma cell migration and invasion, and inhibited EMT by increasing E-cadherin and decreasing N-cadherin and Zeb1.
- Crk was confirmed as a direct target of miR-194-5p.
- miR-194-5p overexpression deactivated the HGF/c-Met/Scr signaling pathway via targeting Crk.
- Crk knockdown reversed the effects of miR-194-5p inhibition on cell invasion, migration, and EMT.
Conclusions:
- miR-194-5p inhibits nephroblastoma cell metastasis and EMT in Wilms tumor progression by targeting Crk.
- The miR-194-5p/Crk axis modulates the HGF/c-Met/Scr signaling pathway, influencing WT cell behavior.
- miR-194-5p represents a promising therapeutic target for Wilms tumor, particularly for preventing metastasis and EMT.
Related Concept Videos
02:30Metastasis
Epithelial-to-Mesenchymal Transition
The epithelial-to-mesenchymal transition or EMT is a developmental process commonly observed in wound healing, embryogenesis, and cancer metastasis. EMT is induced by transforming growth factor-beta (TGF-β) or receptor tyrosine kinase (RTK) ligands, which further...
02:38Mitogens and the Cell Cycle