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1Third Military Medical University, Chongqing, People's Republic of China.
Abstract:
Apurinic/apyrimidinic endonuclease1 (APE1), which has the dual functions of DNA repair and redox regulation, is considered to be a promising potential target in cancer treatment. Microarray and qRT-PCR were used to confirm the change of miRNA followed by analysis with comprehensive bioinformatics-based analysis. Both microarray and qRT-PCR demonstrated that 13 microRNAs (miRNAs) were significantly changed (>2-fold) in APE1 knockdown HOS cells; seven of them (hsa-miR-451, hsa-miR-1290, hsa-miR-765, hsa-miR-483-5p, hsa-miR-513a-5p, hsa-miR-129-5p and hsa-miR-31) were up-regulated and the other six (hsa-miR-29b, hsa-miR-197, has-let-7b, hsa-miR-324-5p, hsa-let-7i and hsa-miR-484) were down-regulated. Furthermore, pathway analysis showed that these miRNAs and their target genes affected by the expression of APE1 were involved in pathways relating to developmental processes, regulation of cellular processes, cell signaling (such as TGF-β, Wnt, MAPK and the p53 signaling pathway) and cancers. There are putative binding sites of NF-κB, p53, HIF-1α, AP-1, PEBP2, ATF, NF-Y, Pax-2,CREB and c-Myb in the promoters of several down regulated miRNAs, indicating that APE1 may regulate miRNAs via transcription factors. Our data suggest that our understanding of the biological functions of APE1 will inevitably expand due to the novel pathways that APE1 uses to regulate gene expression through miRNAs.
Insights
Apurinic/apyrimidinic endonuclease1 (APE1) regulates gene expression through microRNAs (miRNAs). APE1 knockdown significantly altered 13 miRNAs, impacting cancer-related pathways and suggesting novel regulatory mechanisms.
Area of Science:
- Molecular Biology
- Cancer Research
- Genetics
Background:
- Apurinic/apyrimidinic endonuclease1 (APE1) is a key protein with dual roles in DNA repair and redox regulation.
- APE1 is recognized as a potential therapeutic target in cancer treatment.
- Understanding APE1's regulatory mechanisms is crucial for developing novel cancer therapies.
Purpose of the Study:
- To investigate the role of APE1 in regulating microRNA (miRNA) expression.
- To identify specific miRNAs affected by APE1 knockdown.
- To explore the downstream pathways influenced by APE1-mediated miRNA regulation.
Main Methods:
- Microarray analysis to screen for differentially expressed miRNAs.
- Quantitative real-time PCR (qRT-PCR) to validate miRNA expression changes.
- Bioinformatic analysis to predict target genes and associated signaling pathways.
Main Results:
- APE1 knockdown in HOS cells resulted in significant changes (>2-fold) in 13 miRNAs.
- Seven miRNAs were upregulated, and six were downregulated.
- Affected miRNAs and their targets are involved in developmental, cellular regulation, cell signaling (TGF-β, Wnt, MAPK, p53), and cancer pathways.
- Putative transcription factor binding sites (NF-κB, p53, HIF-1α, etc.) were identified in the promoters of downregulated miRNAs.
Conclusions:
- APE1 regulates gene expression through the modulation of miRNA profiles.
- APE1 influences critical cancer-related signaling pathways via miRNA regulation.
- These findings reveal novel mechanisms of APE1 function and expand our understanding of its role in gene expression regulation.
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