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Published on: March 17, 2018
MiR-214-3p targets Ras-related protein 14 (RAB14) to inhibit cellular migration and invasion in esophageal Cancer
Pornima Phatak1,2,3, Whitney M Burrows4, Timothy Michael Creed5
1Birmingham Veterans Affairs Medical Center, Birmingham, AL, USA. pornima.phatak@va.gov.
Background:
MicroRNA (miR)-214-3p is emerging as an important tumor suppressor in esophageal cancer. In this study, we examined the interaction between miR-214-3p and RAB14, a membrane trafficking protein shown to exert oncogenic functions in other malignancies, in esophageal cancer cells.
Methods:
Studies were performed in a human esophageal epithelial cell line and a panel of esophageal cancer cell lines, as well in human specimens. MiR-214-3p expression was measured by digital PCR. Biotinylated RNA pull-down and luciferase reporter assays assessed binding. The xCELLigence RTCA system measured cell migration and invasion in real time. A lentiviral expression vector was used to create an esophageal cancer cell line stably expressing miR-214-3p.
Results:
MiR-214-3p expression was decreased in esophageal cancer cell lines and human specimens compared to non-malignant controls. RAB14 mRNA stability and protein expression were decreased following miR-214-3p overexpression. Binding between miR-214-3p and RAB14 mRNA was observed. Either forced expression of miR-214-3p or RAB14 silencing led to a marked decrease in cellular migration and invasion. Esophageal cancer cells stably expressing miR-214-3p demonstrated decreased growth in a subcutaneous murine model.
Conclusions:
These results further support the tumor-suppressive role of miR-214-3p in esophageal cancer cells by demonstrating its ability to regulate RAB14 expression.
Insights
MicroRNA-214-3p acts as a tumor suppressor in esophageal cancer by regulating RAB14. Overexpressing miR-214-3p or silencing RAB14 inhibits cancer cell migration and invasion.
Area of Science:
- Oncology
- Molecular Biology
- Gene Regulation
Background:
- MicroRNA (miR)-214-3p is recognized as a potential tumor suppressor in esophageal cancer.
- RAB14, a membrane trafficking protein, exhibits oncogenic properties in various cancers.
Purpose of the Study:
- To investigate the interaction between miR-214-3p and RAB14 in esophageal cancer.
- To elucidate the role of miR-214-3p in regulating RAB14 expression and its impact on esophageal cancer progression.
Main Methods:
- Expression analysis of miR-214-3p in esophageal cancer cell lines and human specimens using digital PCR.
- Assessment of miR-214-3p and RAB14 mRNA/protein interaction via biotinylated RNA pull-down and luciferase reporter assays.
- Evaluation of cell migration and invasion using the xCELLigence RTCA system and in vivo tumor growth in a murine model.
Main Results:
- Esophageal cancer tissues and cell lines showed reduced miR-214-3p expression compared to normal controls.
- Overexpression of miR-214-3p led to decreased RAB14 mRNA stability and protein levels, confirming their binding.
- Both miR-214-3p overexpression and RAB14 silencing significantly reduced esophageal cancer cell migration and invasion.
- Stable miR-214-3p expression in esophageal cancer cells resulted in diminished tumor growth in vivo.
Conclusions:
- miR-214-3p functions as a tumor suppressor in esophageal cancer.
- The tumor-suppressive activity of miR-214-3p is mediated through the regulation of RAB14 expression.
- Targeting the miR-214-3p/RAB14 axis may offer therapeutic strategies for esophageal cancer.
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