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Modeling Oral-Esophageal Squamous Cell Carcinoma in 3D Organoids
Published on: December 23, 2022
MicroRNA-transcription factor network analysis reveals miRNAs cooperatively suppress RORA in oral squamous cell
Xueqing Zheng1,2, Kejing Wu3, Shengjie Liao3,4
1The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei_MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, China.
Abstract:
Oral squamous cell carcinoma (OSCC) represents over 90% of oral cancer incidence, while its mechanisms of tumorigenesis remain poorly characterized. In this study, we applied RNA-seq and microRNA-seq methodologies in four pairs of cancer and adjacent normal tissues to profile the contribution of miRNAs to tumorigenesis-altered functional pathways by constructing a comprehensive miRNA-mediated mRNA regulatory network. There were 213 differentially expressed (DE) miRNAs and 2172 DE mRNAs with the involvement of negative miRNA-mRNA interactions identified by at least two pairs of cancerous tissues. GO analysis revealed that the upregulated microRNAs significantly contributed to a global down-regulation of a number of transcription factors (TFs) in OSCC. Among the negative regulatory networks between the selected miRNAs (133) and TFs (167), circadian rhythm genes (RORA, RORB, RORC, and CLOCK) simultaneously regulated by multiple microRNAs were of particular interest. For instance, RORA transcript was predicted to be targeted by 25 co-upregulated miRNAs, of which, miR-503-5p, miR-450b-5p, miR-27a-3p, miR-181a-5p and miR-183-5p were further validated to directly target RORA, resulting in a stronger effect on RORA suppression together. In addition, we showed that the mRNA and protein expression levels of RORα were significantly decreased in most OSCC samples, associated with advanced clinical stage and poor prognosis. RORα significantly suppressed the proliferation of OSCC cells in vitro and in vivo. Attenuated RORα decreased p53 protein expression and suppressed p53 phosphorylation activity. Altogether, our results strongly suggest the importance of the role of miRNAs in regulating the activity of circadian rhythm-related TFs network during OSCC tumorigenesis, and provide further clues to understand the clinical link between circadian rhythm and cancer therapy.
Insights
MicroRNAs regulate circadian rhythm genes in oral squamous cell carcinoma (OSCC), impacting tumor growth. Decreased RORα expression correlates with poor prognosis and suppressed cancer cell proliferation.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Oral squamous cell carcinoma (OSCC) is the most common oral cancer, but its tumorigenesis mechanisms are not fully understood.
- MicroRNAs (miRNAs) play crucial roles in gene regulation and cancer development.
- Circadian rhythm genes are implicated in various cancers, but their specific role in OSCC is unclear.
Purpose of the Study:
- To investigate the role of miRNAs in OSCC tumorigenesis by constructing a miRNA-mediated mRNA regulatory network.
- To identify key miRNA-target interactions and functional pathways altered in OSCC.
- To explore the involvement of circadian rhythm-related genes in OSCC development and prognosis.
Main Methods:
- RNA sequencing (RNA-seq) and microRNA sequencing (miRNA-seq) were performed on four pairs of OSCC and adjacent normal tissues.
- A comprehensive miRNA-mRNA regulatory network was constructed to identify differentially expressed miRNAs and mRNAs.
- Gene Ontology (GO) analysis was used to determine the functional impact of miRNAs on transcription factors (TFs).
- Specific miRNA-TF interactions, particularly involving circadian rhythm genes like RORA, were validated.
- In vitro and in vivo assays were conducted to assess the functional role of RORα in OSCC proliferation and its effect on p53 signaling.
Main Results:
- 213 differentially expressed miRNAs and 2172 differentially expressed mRNAs were identified, with significant negative miRNA-mRNA interactions.
- Upregulated miRNAs in OSCC led to the down-regulation of several transcription factors.
- Circadian rhythm genes, including RORA, were found to be regulated by multiple miRNAs.
- Specific miRNAs were validated to directly target and suppress RORA expression.
- Decreased RORα expression in OSCC tissues correlated with advanced clinical stage and poor prognosis.
- RORα suppressed OSCC cell proliferation and modulated p53 protein expression and phosphorylation.
Conclusions:
- miRNAs play a significant role in regulating circadian rhythm-related transcription factors during OSCC tumorigenesis.
- The identified miRNA-TF regulatory network provides insights into OSCC development.
- RORα acts as a tumor suppressor in OSCC, and its dysregulation by miRNAs contributes to cancer progression.
- These findings suggest potential therapeutic strategies targeting the circadian rhythm pathway in OSCC.
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