Dysregulation of miR-1-3p: An Early Event in Colitis-Associated Dysplasia
Abstract
:1. Introduction
2. Results
2.1. miRNA Signature Differentiates Inflamed Non-Neoplastic Mucosa from Flat and Polypoid Dysplasia
2.2. Genes Differentially Expressed in Colitis-Associated Colonic Dysplasias
2.3. miR Targets Are Associated with the Progression of Colitis-Associated Carcinogenesis
2.4. miR-30c, miR-145a and miR-1 Are the Hub miRs of the Early Detection Signature
2.5. Protein–Protein Interactions Categorize the Selected Targets into Five Functional Clusters
2.6. Biological Function of miR-1 In Vitro
2.7. miR-1 Induces Apoptosis in HCT116 and RKO Human Colon Carcinoma Cells
2.8. miR-1 Induces Cell Cycle Arrest in G0/G1
2.9. miR-1 Increases the Number of Cells in Sub-G1
2.10. miR-1 Targets the 3′UTR of Cdk6 in Colon Carcinoma Cells
3. Discussion
4. Materials and Methods
4.1. Experimental Design
4.2. Laser Microdissection and RNA Extraction
4.3. nCounter® Mouse miRNA Expression Panel
4.4. Global Gene Expression Microarray
4.5. Validation of miRNA Signature by RT-qPCR
4.6. Target Prediction and Pathway Enrichment Analysis
4.7. Analysis of Centrality of the miR Signature and Construction of an Interaction Network
4.8. Construction of 3′-UTR Reporter Plasmids and the Cloning Method
4.9. Cotransfection of Plasmid DNA, miRNA Mimics and pRL-TK Vector
4.10. Flow Cytometry Analyses for Apoptosis and Cell Cycle
4.11. Cdk6 Expression in Human Colorectal Cancer Cells by RT-qPCR
4.12. Statistical Analyses
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Fragoso, M.F.; Fernandez, G.J.; Vanderveer, L.; Cooper, H.S.; Slifker, M.; Clapper, M.L. Dysregulation of miR-1-3p: An Early Event in Colitis-Associated Dysplasia. Int. J. Mol. Sci. 2022, 23, 13024. https://doi.org/10.3390/ijms232113024
Fragoso MF, Fernandez GJ, Vanderveer L, Cooper HS, Slifker M, Clapper ML. Dysregulation of miR-1-3p: An Early Event in Colitis-Associated Dysplasia. International Journal of Molecular Sciences. 2022; 23(21):13024. https://doi.org/10.3390/ijms232113024
Chicago/Turabian StyleFragoso, Mariana F., Geysson J. Fernandez, Lisa Vanderveer, Harry S. Cooper, Michael Slifker, and Margie L. Clapper. 2022. "Dysregulation of miR-1-3p: An Early Event in Colitis-Associated Dysplasia" International Journal of Molecular Sciences 23, no. 21: 13024. https://doi.org/10.3390/ijms232113024