Down-Regulation of miR-129-5p and the let-7 Family in Neuroendocrine Tumors and Metastases Leads to Up-Regulation of Their Targets Egr1, G3bp1, Hmga2 and Bach1
Abstract
:1. Introduction
2. Experimental
2.1. Clinical Samples
2.2. Cell Culture
2.3. RNA Extraction
2.4. miRNA Microarray Analyses
2.5. qPCR
2.6. In Situ Hybridization
2.7. Immunohistochemistry
2.8. Transfection Studies and Cell Growth Analyses
2.9. mRNA Microarray Analysis
2.10. Evaluating Global Down-Regulation of microRNA Target Genes
2.12. Gene Set Enrichment Analysis
2.13. 3'UTR Luciferase Assay
2.14. Western Blotting
2.15. Statistical Analyses
3. Results
3.1. miR-129-5p is Down-Regulated in Neuroendocrine Tumors and the let-7 Family is Down-Regulated in Neuroendocrine Tumor Metastases
T + M vs. N | Log FC | FC | p-value |
---|---|---|---|
hsa-miR-129-5p | −1.3 | 0.4 | 0.01 |
hsa-let-7a | −1.7 | 0.3 | 0.002 |
hsa-let-7c | −1.7 | 0.3 | 0.001 |
hsa-let-7b | −1.5 | 0.3 | 0.0003 |
hsa-let-7d | −1.5 | 0.3 | 0,003 |
hsa-let-7e | −1.4 | 0.4 | 0.002 |
hsa-let-7f | −1.4 | 0.4 | 0.003 |
hsa-let-7g | −1.1 | 0.5 | 0.01 |
hsa-let-7i | −1.3 | 0.4 | 0.001 |
M vs. N | Log FC | FC | p-value |
hsa-let-7a | −1.7 | 0.3 | 0.002 |
hsa-let-7c | −1.7 | 0.3 | 0.001 |
hsa-let-7b | −1.5 | 0.3 | 0.0003 |
hsa-let-7d | −1.5 | 0.3 | 0.003 |
hsa-let-7e | −1.4 | 0.4 | 0.002 |
hsa-let-7f | −1.4 | 0.4 | 0.003 |
hsa-let-7g | −1.1 | 0.5 | 0.01 |
hsa-let-7i | −1.3 | 0.4 | 0.001 |
M vs. T | Log FC | FC | p-value |
hsa-let-7a | −0.03 | 0.8 | 0.3 |
hsa-let-7c | −0.02 | 1.0 | 0.9 |
hsa-let-7b | −0.3 | 0.8 | 0.3 |
hsa-let-7d | −0.2 | 0.9 | 0.6 |
hsa-let-7e | −0.1 | 0.9 | 0.6 |
hsa-let-7f | −0.1 | 0.9 | 0.8 |
hsa-let-7g | −0.1 | 0.9 | 0.8 |
hsa-let-7i | −0.02 | 1.0 | 0.9 |
3.2. miR-129-5p Inhibits Growth of the Carcinoid Cell Lines NCI-H727 and CNDT2
3.3. miR-129-5p Targets EGR1 and G3BP1 in Carcinoid Cell Lines NCI-H727 and CNDT2
3.4. EGR1 and G3BP1 are Up-Regulated in Neuroendocrine Tumors and Promote Growth in Carcinoid Cell Lines
3.5. The let-7 Family is Involved in the Neuroendocrine Tumor Metastatic Process by Targeting HMGA2 and BACH1
4. Discussion
5. Conclusions
Supplementary Files
Supplementary File 1Acknowledgments
Author Contributions
Conflicts of Interest
References
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Døssing, K.B.V.; Binderup, T.; Kaczkowski, B.; Jacobsen, A.; Rossing, M.; Winther, O.; Federspiel, B.; Knigge, U.; Kjær, A.; Friis-Hansen, L. Down-Regulation of miR-129-5p and the let-7 Family in Neuroendocrine Tumors and Metastases Leads to Up-Regulation of Their Targets Egr1, G3bp1, Hmga2 and Bach1. Genes 2015, 6, 1-21. https://doi.org/10.3390/genes6010001
Døssing KBV, Binderup T, Kaczkowski B, Jacobsen A, Rossing M, Winther O, Federspiel B, Knigge U, Kjær A, Friis-Hansen L. Down-Regulation of miR-129-5p and the let-7 Family in Neuroendocrine Tumors and Metastases Leads to Up-Regulation of Their Targets Egr1, G3bp1, Hmga2 and Bach1. Genes. 2015; 6(1):1-21. https://doi.org/10.3390/genes6010001
Chicago/Turabian StyleDøssing, Kristina B. V., Tina Binderup, Bogumil Kaczkowski, Anders Jacobsen, Maria Rossing, Ole Winther, Birgitte Federspiel, Ulrich Knigge, Andreas Kjær, and Lennart Friis-Hansen. 2015. "Down-Regulation of miR-129-5p and the let-7 Family in Neuroendocrine Tumors and Metastases Leads to Up-Regulation of Their Targets Egr1, G3bp1, Hmga2 and Bach1" Genes 6, no. 1: 1-21. https://doi.org/10.3390/genes6010001