The Role of miR-375-3p and miR-200b-3p in Gastrointestinal Stromal Tumors
Abstract
:1. Introduction
2. Results
2.1. Overexpression of miR-375-3p and miR-200b-3p Alters the Expression of Their Putative Target Genes
2.2. miR-375-3p and miR-200b-3p Directly Regulate Their Predicted Targets KIT and EGFR
2.3. miR-375-3p Reduced Cell Viability and Proliferation
2.4. miR-375-3p and miR-200b-3p Reduced Cell Migration Rate
2.5. miR-375-3p and miR-200b-3p Did Not Affect Cell Apoptosis
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. miRNA Selection and Target Prediction
4.3. Cell Transfection
4.4. Quantitative Reverse Transcription PCR
4.5. Western Blot
4.6. Luciferase Reporter Assay
4.7. MTT Assay
4.8. Apoptosis Assay
4.9. Wound Healing Assay
4.10. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
3′UTR | three prime untranslated region |
ATCC | American type culture collection |
DMSO | dimethyl sulfoxide |
EGFR | epidermal growth factor receptor |
ETV1 | ETS transcription factor 1 |
FITC | fluorescein isothiocyanate |
GAPDH | glyceraldehyde 3-phosphate dehydrogenase |
GIST | gastrointestinal stromal tumor |
JAK | Janus kinase |
MAPK | mitogen-activated protein kin |
miRNA | microRNA |
mTOR | mammalian target of rapamycin |
MTT | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide |
mut | mutant |
NC | negative control |
PDGFRA | platelet-derived growth factor receptor-α |
PI | propidium iodide |
PI3K | phosphatidylinositol-3-kinase |
RIPA | radioimmunoprecipitation assay |
RNA | ribonucleic acid |
RT-qPCR | Reverse transcription quantitative polymerase chain reaction |
RTK | receptor tyrosine kinase |
STAT | signal transducer and activator of transcription |
wt | wild-type |
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Gyvyte, U.; Lukosevicius, R.; Inciuraite, R.; Streleckiene, G.; Gudoityte, G.; Bekampyte, J.; Valentini, S.; Salteniene, V.; Ruzgys, P.; Satkauskas, S.; et al. The Role of miR-375-3p and miR-200b-3p in Gastrointestinal Stromal Tumors. Int. J. Mol. Sci. 2020, 21, 5151. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21145151
Gyvyte U, Lukosevicius R, Inciuraite R, Streleckiene G, Gudoityte G, Bekampyte J, Valentini S, Salteniene V, Ruzgys P, Satkauskas S, et al. The Role of miR-375-3p and miR-200b-3p in Gastrointestinal Stromal Tumors. International Journal of Molecular Sciences. 2020; 21(14):5151. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21145151
Chicago/Turabian StyleGyvyte, Ugne, Rokas Lukosevicius, Ruta Inciuraite, Greta Streleckiene, Greta Gudoityte, Justina Bekampyte, Serena Valentini, Violeta Salteniene, Paulius Ruzgys, Saulius Satkauskas, and et al. 2020. "The Role of miR-375-3p and miR-200b-3p in Gastrointestinal Stromal Tumors" International Journal of Molecular Sciences 21, no. 14: 5151. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21145151