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. 2022 Jun 20;24(2):270.
doi: 10.3892/ol.2022.13390. eCollection 2022 Aug.

Identification of circular RNA_0000919 as a potential diagnostic and prognostic biomarker of tongue squamous cell carcinoma using circular RNA microarray and reverse transcription-quantitative PCR analyses

Hongli Liu  1 , Qi Li  1 , Han Qi  1 , Fengzhi Du  1 , Yanli Qiu  1
Affiliations

Identification of circular RNA_0000919 as a potential diagnostic and prognostic biomarker of tongue squamous cell carcinoma using circular RNA microarray and reverse transcription-quantitative PCR analyses

Hongli Liu et al. Oncol Lett. .

Abstract

The present study aimed to identify differentially expressed (DE) circular RNAs (circRNAs/circs) using microarray analysis and to further explore the clinical significance of 10 candidate DEcircRNAs in patients with tongue squamous cell carcinoma (TSCC). A total of 60 patients with TSCC who underwent surgery were enrolled and five pairs of TSCC and adjacent (Ctrl) tissues were used for circRNA microarray analysis. Subsequently, the top five upregulated and downregulated DEcircRNAs were detected by reverse transcription-quantitative PCR (RT-qPCR) analysis in 60 pairs of tumor and Ctrl tissues, and their association with tumor features and overall survival (OS) was further analyzed. circRNA expression was used to differentiate TSCC from Ctrl tissues by principal component and heatmap analyses. A total of 134 upregulated and 67 downregulated DEcircRNAs were identified in TSCC tissues compared with Ctrl tissues. The DEcircRNAs were enriched in oncogenic signaling, including the 'Wnt signaling pathway' and the 'MAPK signaling pathway'. The majority of DEcircRNAs exhibited several target microRNAs (miRNAs) in regulatory network analysis. These findings were validated by RT-qPCR analysis and the results demonstrated that the expression levels of 9/10 selected candidate DEcircRNAs (circ_0020048, circ_0000919, circ_0004525, circ_0002113, circ_0004029, circ_0004503, circ_0008752, circ_0002300 and circ_0001811) were dysregulated in TSCC tissues compared with Ctrl tissues. The expression levels of five DEcircRNAs (circ_0004503, circ_0008752, circ_0002300, circ_0020048 and circ_0000919) were associated with pathological grade or tumor clinical stage. Notably, only the expression levels of one DEcircRNA (circ_0000919) were associated with decreased OS. In conclusion, the present study indicated aberrant circRNA expression and potential circRNA-miRNA interactions in TSCC and identified circ_0000919 as a diagnostic and prognostic biomarker for TSCC management.

Keywords: OS; RT-qPCR; TSCC; circRNA; microarray and bioinformatics analyses.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1.
Figure 1.
PCA and heatmap analysis. (A) PCA and (B) heatmap analysis of circRNA expression profile in tumor and paired Ctrl tissues derived from patients with TSCC. circRNA, circular RNA; Ctrl, paired adjacent normal tissues; Dim, dimension; PCA, principal component analysis; TSCC, tongue squamous cell carcinoma.
Figure 2.
Figure 2.
Volcano plot of the circRNA expression profile noted in tongue squamous cell carcinoma and paired adjacent tissues. circRNA, circular RNA; DEcircRNA, differentially expressed circRNA; DOWN, downregulated; Padj, adjusted P-value; UP, upregulated.
Figure 3.
Figure 3.
Enrichment analysis. GO enrichment analysis of DEcircRNAs based on (A) the gene function and (B) the target miRNA in patients with TSCC. KEGG enrichment analysis for DEcircRNAs based on (C) the gene function and (D) the target miRNA in patients with TSCC. circRNA, circular RNA; DE, differentially expressed; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; miRNA; microRNA; TSCC; tongue squamous cell carcinoma.
Figure 4.
Figure 4.
Network of circRNA-miRNA interactions. Interaction of top 10 candidate differentially expressed circRNAs with their potential target miRNAs in tongue squamous cell carcinoma. The heatmap is a graphical presentation of the data composed of grids of colors and clusters on both rows and columns and provides an overview of the numerical differences. Squares indicate the circRNAs and circles indicate miRNAs. circRNAs in red are upregulated circRNAs; circRNAs in blue are downregulated circRNAs; miRNAs in green are the potential targets of circRNAs. circRNA/circ, circular RNA; miRNA/miR, microRNA.
Figure 5.
Figure 5.
Validation of candidate differentially expressed circRNA expression levels in patients with TSCC. Comparison of the expression levels of (A) circ_0020048, (B) circ_0000919, (C) circ_0004525, (D) circ_0066887, (E) circ_0002113, (F) circ_0004029, (G) circ_0004503, (H) circ_0008752, (I) circ_0002300 and (J) circ_0001811 between 60 TSCC and adjacent tissues. circRNA/circ, circular RNA; TSCC, tongue squamous cell carcinoma.
Figure 6.
Figure 6.
Association between candidate differentially expressed circRNAs and prognosis of patients with TSCC. Association between the expression levels of (A) circ_0020048, (B) circ_000919, (C) circ_0004525, (D) circ_0066887, (E) circ_0002113, (F) circ_0004029, (G) circ_0004503, (H) circ_0008752, (I) circ_0002300 and (J) circ_0001811 in tumors derived from 60 patients with TSCC and overall survival. 95% CI, 95% confidence interval; circRNA/circ, circular RNA; HR, hazard ratio; OS, overall survival; TSCC, tongue squamous cell carcinoma.
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References

    1. Kim YJ, Kim JH. Increasing incidence and improving survival of oral tongue squamous cell carcinoma. Sci Rep. 2020;10:7877. doi: 10.1038/s41598-020-64748-0. - DOI - PMC - PubMed
    1. Shrestha AD, Vedsted P, Kallestrup P, Neupane D. Prevalence and incidence of oral cancer in low- and middle-income countries: A scoping review. Eur J Cancer Care (Engl) 2020;29:e13207. doi: 10.1111/ecc.13207. - DOI - PubMed
    1. Ohta K, Yoshimura H. Squamous cell carcinoma of the dorsal tongue. CMAJ. 2019;191:E1310. doi: 10.1503/cmaj.190677. - DOI - PMC - PubMed
    1. Machiels JP, René Leemans C, Golusinski W, Grau C, Licitra L, Gregoire V. secretariat@ehns.orgclinicalguidelines@esmo.orginfo@estro.orgSquamous cell carcinoma of the oral cavity, larynx, oropharynx and hypopharynx: EHNS-ESMO-ESTRO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol. 2020;31:1462–1475. doi: 10.1016/j.annonc.2020.07.011. EHNS Executive Board. Electronic address. ESMO Guidelines Committee. Electronic address. ESTRO Executive Board. Electronic address. - DOI - PubMed
    1. Okubo M, Iwai T, Nakashima H, Koizumi T, Oguri S, Hirota M, Mitsudo K, Tohnai I. Squamous Cell Carcinoma of the Tongue Dorsum: Incidence and Treatment Considerations. Indian J Otolaryngol Head Neck Surg. 2017;69:6–10. doi: 10.1007/s12070-016-0979-z. - DOI - PMC - PubMed