miR-20b and miR-451a Are Involved in Gastric Carcinogenesis through the PI3K/AKT/mTOR Signaling Pathway: Data from Gastric Cancer Patients, Cell Lines and Ins-Gas Mouse Model
Abstract
:1. Introduction
2. Results
2.1. Aberrant Expression of miR-20b and miR-451a In Vivo
2.2. Aberrant Expression of miR-20b and miR-451a in Vitro
2.3. Inhibition of miR-20b Reduced Cell Viability and Proliferation
2.4. Inhibition of miR-20b and Overexpression of miR-451a Dramatically Reduced Colony Formation Rate
2.5. Inhibition of miR-20b Increased Cell Apoptosis Rate
2.6. miR-20b and miR-451a Have No Effect In Vitro On Cell Migration Processes
2.7. PTEN and TXNIP Are Direct Targets of miR-20b
2.8. miR-451a Directly Regulate the Expression of CAV1 and TSC1
3. Discussion
4. Materials and Methods
4.1. Human Tissue Samples and Cell Lines
4.2. Cell Transfection
4.3. Target Prediction
4.4. qRT-PCR for miR-20b and miR-451a Expression Level
4.5. Western Blot
4.6. Luciferase Reporter Assay
4.7. MTT Assay
4.8. Colony Formation Assay
4.9. Apoptosis Assay
4.10. Wound Healing Assay
4.11. Tumorigenicity of miR-20b and miR-451a in INS-GAS Mouse Model
4.12. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
3′ UTR | three prime untranslated region |
ATCC | American type culture collection |
CAV1 | caveolin-1 |
DMSO | dimethyl sulfoxide |
FBS | fetal bovine serum |
FITC | fluorescein isothiocyanate |
GAPDH | glyceraldehyde 3-phosphate dehydrogenase |
GC | gastric cancer |
HER | human epidermal growth factor receptor |
INS-GAS | insulin-gastrin |
IRF1 | interferon regulatory factor 1 |
miRNA | microRNA |
mTOR | mammalian target of rapamycin |
MTT | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide |
mut | mutant |
OD | optical density |
PI | propidium iodide |
PI3K | phosphatidylinositol-3-kinase |
PTEN | phosphatase and tensin homologue |
RIPA | radioimmunoprecipitation assay |
RISC | RNA-induced silencing complex |
RNA | ribonucleic acid |
TSC1 | tuberous sclerosis 1 |
TXNIP | thioredoxin-interacting protein |
wt | wild-type |
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Streleckiene, G.; Inciuraite, R.; Juzenas, S.; Salteniene, V.; Steponaitiene, R.; Gyvyte, U.; Kiudelis, G.; Leja, M.; Ruzgys, P.; Satkauskas, S.; et al. miR-20b and miR-451a Are Involved in Gastric Carcinogenesis through the PI3K/AKT/mTOR Signaling Pathway: Data from Gastric Cancer Patients, Cell Lines and Ins-Gas Mouse Model. Int. J. Mol. Sci. 2020, 21, 877. https://doi.org/10.3390/ijms21030877
Streleckiene G, Inciuraite R, Juzenas S, Salteniene V, Steponaitiene R, Gyvyte U, Kiudelis G, Leja M, Ruzgys P, Satkauskas S, et al. miR-20b and miR-451a Are Involved in Gastric Carcinogenesis through the PI3K/AKT/mTOR Signaling Pathway: Data from Gastric Cancer Patients, Cell Lines and Ins-Gas Mouse Model. International Journal of Molecular Sciences. 2020; 21(3):877. https://doi.org/10.3390/ijms21030877
Chicago/Turabian StyleStreleckiene, Greta, Ruta Inciuraite, Simonas Juzenas, Violeta Salteniene, Ruta Steponaitiene, Ugne Gyvyte, Gediminas Kiudelis, Marcis Leja, Paulius Ruzgys, Saulius Satkauskas, and et al. 2020. "miR-20b and miR-451a Are Involved in Gastric Carcinogenesis through the PI3K/AKT/mTOR Signaling Pathway: Data from Gastric Cancer Patients, Cell Lines and Ins-Gas Mouse Model" International Journal of Molecular Sciences 21, no. 3: 877. https://doi.org/10.3390/ijms21030877