MiRNA-424-5p Suppresses Proliferation, Migration, and Invasion of Clear Cell Renal Cell Carcinoma and Attenuates Expression of O-GlcNAc-Transferase

• Published in: Cancers 2021-10, Vol.13 (20), p.5160
• Main Authors: Kalantzakos, Thomas J., Sullivan, Travis B., Gloria, Thales, Canes, David, Moinzadeh, Alireza, Rieger-Christ, Kimberly M.
• Format: Article
• Language: English
• Subjects: Apoptosis; Biomarkers; Cell culture; Cell growth; Cell migration; Cell proliferation; Cell viability; Clear cell-type renal cell carcinoma; DNA methylation; Epigenetics; Gene expression; Kidney cancer; Medical prognosis; Metastases; Metastasis; miRNA; Phenotypes; Post-transcription; Proteins; Transfection; Tumors; Western blotting
• ISSN: 2072-6694; 2072-6694
• DOI: 10.3390/cancers13205160

MicroRNAs (miRNAs) are non-coding post-transcriptional regulators of gene expression that are dysregulated in clear cell renal cell carcinoma (ccRCC) and play an important role in tumor progression. Our prior work identified a subset of miRNAs in pT1 ccRCC tumors, including miR-424-5p, that are associated with an aggressive phenotype. We investigate the impact of this dysregulated miRNA and its protein target O-GlcNAc-transferase (OGT) to better understand the mechanisms behind aggressive stage I ccRCC. The ccRCC cell lines 786-O and Caki-1 were used to assess the impact of miR-424-5p and OGT. Cells were transfected with pre-miR-424-5p, a lentiviral anti-OGT shRNA, or were treated with the demethylating agent 5-Aza-2′-deoxycytidine. Cell proliferation was measured via MT cell viability assay. Cell migration and invasion were analyzed using Transwell assays. The expression of miR-424-5p was determined through qRT-PCR, while OGT protein expression was evaluated through Western blotting. The interaction between miR-424-5p and OGT was confirmed via luciferase reporter assay. The transfection of ccRCC cells with pre-miR-424-5p or anti-OGT shRNA significantly inhibited cell proliferation, migration, and OGT expression, while miR-424-5p also attenuated cell invasion. Addition of the demethylating agent significantly reduced cell proliferation, migration, invasion, and OGT expression, while significantly increasing the expression of miR-424-5p. Altogether, these findings suggest that epigenetic downregulation of miR-424-5p, which in turn augments OGT expression, contributes to the creation of aggressive forms of stage I ccRCC.