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YTHDF2-regulated hsa_circ_0005882 functions as a sponge for miR-654-3p to suppress nasopharyngeal carcinoma
Xu Wang1,2, Aiyu Ma1,2, Lu Lu1
1Department of Pathology, The First Affiliated Hospital of Guilin Medical University, Guilin, Guangxi, China.
Abstract:
Circular RNAs (circRNAs) are a subclass of non-coding RNAs, playing an important regulatory role in tumor progression. Accumulating evidence has revealed that circRNAs can be regulated by m6A machinery, influencing their expression and biological functions. However, the functions and mechanisms of m6A-mediated circRNAs in nasopharyngeal carcinoma (NPC) remain to be elucidated. In this study, the sequence of hsa_circ_0005882 (circ_0005882) was determined by Sanger sequencing, and its localization in both the cytoplasm and nucleus was confirmed by fluorescence in situ hybridization. Furthermore, m6A RNA immunoprecipitation followed by qPCR (MeRIP-qPCR) and the single-base elongation- and ligation-based qPCR amplification (SELECT) assays revealed that circ_0005882 may harbor m6A modifications. Overexpression of circ_0005882 significantly suppressed NPC cell proliferation. Mechanistically, the m6A reader YTHDF2 binds to circ_0005882 to promote its degradation, thereby reducing circ_0005882 stability. Additionally, circ_0005882 functions as a sponge for miR-654-3p, contributing to the inhibition of NPC proliferation. These findings collectively demonstrate that circ_0005882, which is negatively regulated by YTHDF2, functions as a tumor suppressor by sponging miR-654-3p to inhibit NPC cell proliferation, unveiling a novel regulatory model centered on the YTHDF2/circ_0005882/miR-654-3p axis in NPC pathogenesis.
Insights
This study reveals that circ_0005882 acts as a tumor suppressor in nasopharyngeal carcinoma (NPC) by sponging miR-654-3p. Its stability is negatively regulated by YTHDF2, highlighting a new YTHDF2/circ_0005882/miR-654-3p axis in NPC.
Area of Science:
- Molecular Biology
- Oncology
- RNA Biology
Background:
- Circular RNAs (circRNAs) are key regulators in tumor progression.
- The role of m6A-modified circRNAs in nasopharyngeal carcinoma (NPC) is not fully understood.
- Investigating m6A-mediated circRNAs offers potential therapeutic targets for NPC.
Purpose of the Study:
- To elucidate the function and mechanism of m6A-modified circRNAs in NPC.
- To identify specific circRNAs involved in NPC pathogenesis.
- To explore the regulatory network of circRNAs, m6A machinery, and microRNAs in NPC.
Main Methods:
- Sanger sequencing to determine circRNA sequence.
- Fluorescence in situ hybridization for subcellular localization.
- m6A RNA immunoprecipitation followed by qPCR (MeRIP-qPCR) and SELECT assays to detect m6A modification.
- Cell proliferation assays.
- Western blotting and quantitative PCR to analyze protein and RNA levels.
Main Results:
- hsa_circ_0005882 (circ_0005882) was identified and confirmed to be present in both cytoplasm and nucleus.
- circ_0005882 harbors m6A modifications and its overexpression suppresses NPC cell proliferation.
- The m6A reader YTHDF2 binds to circ_0005882, promoting its degradation and reducing its stability.
- circ_0005882 acts as a molecular sponge for miR-654-3p, inhibiting NPC proliferation.
Conclusions:
- circ_0005882 functions as a tumor suppressor in NPC.
- YTHDF2 negatively regulates circ_0005882 stability.
- The YTHDF2/circ_0005882/miR-654-3p axis represents a novel regulatory mechanism in NPC pathogenesis.
