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Navigating The CeRNA Axis by Bioinformatics and Experimental Validation: Identification of ADIRF-AS1, miR-191-5p, and
Azadeh Rezaei1, Rahim Rostami1, Shahram Teimourian2
1Biochemistry Department of Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran.
Background:
Endometrial carcinoma (EC) is a significant gynecologic malignancy. Investigating competing endogenous RNA (ceRNA) networks, including long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and messenger RNAs (mRNAs), offers insights into EC's molecular intricacies and may improve therapeutic and diagnostic strategies. This study focuses on the ceRNA axis, particularly the interactions of lncRNA-miRNA-mRNA-Gene, using TCGA-UCEC database analysis and experimental validation.
Materials And Methods:
In this case-control study, differentially expressed lncRNAs (DElncRNAs), microRNAs (DEMIs), and genes (DEGs) were identified, highlighting ADIRF-AS1 as a potential target. Validated interactions between ADIRF-AS1, miR-191-5p, and EGR1 were established, with significant ADIRF-AS1/EGR1 correlation. Protein-protein interaction (PPI) analysis identified 21 proteins linked to EGR1, with gene ontology (GO) analysis revealing roles in myeloid cell differentiation. Expression levels of genes, lncRNA, and microRNAs were evaluated by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Western blot analysis was applied for protein evaluation.
Results:
RT-qPCR results showed that the RNA expression levels of ADIRF-AS1 and EGR1 genes in endometrial cancer and hyperplasia samples were significantly lower (P<0.001) than control samples. Also, the expression level of miR-191-5p in endometrial cancer tissues was significantly higher than patients with hyperplasia (P<0.001) and normal samples (P<0.001). Western blot results also showed that the protein level of EGR1 in endometrial cancer samples was significantly lower than control samples (P<0.001).
Conclusion:
Here, we observed an interaction between lncRNA ADIRF-AS1and hsa-miR-191-5p, and also, ADIRFAS1 downstream effects on EGR1 in EC, that seems may be a suggesting therapeutic and diagnostic targets. Further research could explore its clinical relevance in endometrial carcinoma.
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