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Circular RNA expression profiling identifies specific circular RNAs in tongue squamous cell carcinoma.

ABSTRACT: Tongue squamous cell carcinoma (TSCC) is the most frequent type of oral cancer associated with high malignancy. Circular RNAs (circRNAs) are a form of non?coding RNA with stable and conserved expression in mammalian cells. The aim of the present study was to investigate circRNAs expression profiles in TSCC, and examine the roles and potential mechanisms of circRNA?081069 (circ_081069). A high?throughput circRNA microarray analysis of tumor samples and adjacent normal tissues from four patients with TSCC was performed. Bioinformatic analysis was conducted to screen the differentially expressed circRNAs. Reverse transcription?quantitative PCR was performed to confirm the microarray results. A migration assay and proliferation assay were performed to detect the migratory and proliferative ability of TSCC cells. A luciferase assay was conducted to investigate the interaction between circ_081069 and microRNA (miRNA/miR)?665. In total, 335 circRNAs were found to be differentially expressed in tumor tissues. Among them, 59 were upregulated and 276 were downregulated (P<0.05; fold change ?2 or ?0.5). A total of seven circRNAs, including two upregulated and five downregulated circRNAs, were further confirmed using quantitative PCR analysis in the ten paired TSCC tissues and adjacent normal tissues. The present study showed that circRNA_081069 promoted the migratory and proliferative ability of TSCC cells in vitro. Furthermore, the potential circRNA?miRNA interactions were predicted, and the present results identified miR?665 as a miRNA target of circ_081069. The present results suggested that circRNAs may be involved in TSCC development, and understanding the interaction between circ_081069 and miR?665 may facilitate the development of novel diagnostic and therapeutic targets for TSCC.

SUBMITTER: Wei T

PROVIDER: S-EPMC7057816 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature

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Circular RNA expression profiling identifies specific circular RNAs in tongue squamous cell carcinoma.

Wei Tai T Ye Peng P Yu Guang-Yan GY Zhang Zu-Yan ZY

Molecular medicine reports 20200206 4

Tongue squamous cell carcinoma (TSCC) is the most frequent type of oral cancer associated with high malignancy. Circular RNAs (circRNAs) are a form of non‑coding RNA with stable and conserved expression in mammalian cells. The aim of the present study was to investigate circRNAs expression profiles in TSCC, and examine the roles and potential mechanisms of circRNA‑081069 (circ_081069). A high‑throughput circRNA microarray analysis of tumor samples and adjacent normal tissues from four patients w ...[more]

PMID: 32319610

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Project description:Circular RNAs are characterized as a class of covalently closed circular RNA transcripts and are associated with a variety of cellular processes and neurological diseases by sponging microRNAs. Expression profiling of circular RNAs in glaucoma, which is a form of optic neuropathy, has not been performed to date. The most common characteristic of all forms of glaucoma is the loss of retinal ganglion cells. While the pathogenesis of glaucoma is not fully understood, intraocular pressure is unquestionably the only proven modifiable factor which makes chronic ocular hypertension (COH) animals the classical glaucoma models. Based on these findings, we completed the first in-depth study of rat retinal circular RNA expression profiling to identify probable biomarkers for the diagnosis of glaucoma. Two ocular hypertension models were induced by episcleral vein ligation (EVL) and microbead injection in rats. Overall, 15,819 circular RNA were detected. Furthermore, 3,502 differentially expressed circular RNAs verified in both COH rats were identified, of which 691 were upregulated and 2,811 were downregulated. Seven significantly downregulated (both log2FoldChange < -2.5 and adjusted P < 0.001) and seven significantly upregulated (both log2FoldChange > 2.5 and adjusted P < 0.001) circular RNAs were shown. Six target microRNAs aligned with the top 14 circular RNAs were identified. According to the construction of the circular RNA-microRNA network and circBase information, only RNO_CIRCpedia_1775 had the homologous hsa_circ_0023826 in the human genome. The hsa_circ_0023826 and mRNA of the host gene TENM4 (teneurin transmembrane protein 4) were validated in aqueous humor samples of five glaucoma patients and five cataract control patients. The expression of hsa_circ_0023826 showed a significant decrease in glaucoma patients, while TENM4 mRNA showed no significant difference compared to cataract patients (P = 0.024 and P = 0.294, respectively). The results of this study comprehensively characterized the expression profiles of circular RNA in glaucoma-affected eyes, as verified by two different ocular hypertension rat models. Together with the target microRNAs underlying the top differentially expressed circular RNAs, a new target of hsa_circ_0023826 and its host gene TENM4 were identified and further verified in the aqueous humor of glaucoma patients, indicating a promising biomarker for the disease.

Project description:BackgroundCircular RNAs (circRNAs) are involved in the pathogenesis of several diseases. Among oral maxillofacial cancers, oral squamous cell carcinoma (OSCC) has the highest incidence. However, the role of circRNAs in OSCC is still not clear. The aim of our study was to evaluate the circRNA expression profile in OSCC and explore further the potential role of circRNAs in the pathogenesis of OSCC.MethodsCircRNA sequencing was performed in 6 pairs of samples of OSCC and normal oral mucosal tissues. Expression of selected circRNAs was validated by qRT-PCR. GO and KEGG analyses were performed and binding relationships between circRNAs and miRNAs were predicted. The hsa_circ_0001766/miR-877-3p/VEGFA axis was selected to further elucidate its role in OSCC.ResultsWe showed that there were 122 differentially expressed (DE) circRNAs. Eight out of 10 selected circRNAs were validated by qRT-PCR. GO and KEGG analyses indicated that the identified DE circRNAs might be involved in the progression of OSCC. Then, after identification by Sanger sequencing and RNase R assay, the upregulated hsa_circ_0001766 was selected to investigate its potential role in OSCC. Bioinformatics analysis showed that hsa_circ_0001766 might act as a competing endogenous RNA (ceRNA) that sponged miR-877-3p to upregulate VEGFA expression. We selected OSCC cell lines SCC9 and SCC25. PCR results showed that the expression of hsa_circ_0001766 and VEGFA was upregulated in SCC9 and SCC25. Subsequently, using western blot, PCR, CCK8, and colony formation assays, we found that knocking down circRNA0001766 inhibited the expression of VEGFA and the proliferation of OSCC cells. Following this, miR-877-3p inhibitor reversed the inhibitory effect of si-hsa_circ_0001766 on expression of VEGFA and proliferation of OSCC cells.ConclusionsIn conclusion, our study revealed the possible role of circRNAs in the pathogenesis of OSCC, and identified the potential role of the hsa_circ_0001766/miR-877-3p/VEGFA axis in OSCC progression.

Project description:Abnormal expression of non-coding circular RNAs (circRNAs) have been reported in many types of tumors. circRNA have been suggested to be an ideal candidate biomarker for diagnostic and therapeutic implications in cancers. The aim of this study was to assess the circRNA expression profile of laryngeal squamous cell carcinomas (LSCC). The biopsy samples from patients with LSCC were obtained intra-operatively. The circRNA expression was performed using secondary sequencing. Among 10 patients with LSCC, 2 were well differentiated, 3 were moderately differentiated and 5 were adjunctive samples with normal and LSCC tissues. A total of 21,444 distinct circRNA candidates were detected. Among them, we defined the statistical criteria for selecting aberrant-expressed circRNA using a q-value of < 0.001 with a fold change of > 2.0 or < 0.5. A total of 29 circRNA were upregulated and 19 circRNA were downregulated significantly in the LSCC tissues. The intersection of these dysregulated circRNAs of normal-well differentiated set and normal-moderately differentiated set was then assessed to narrow the upregulated and downregulated circRNAs down to 18 and 5 respectively. Furthermore, an association of the circRNA-miRNA-mRNA was investigated, showing that 20 dysregulated circRNA successfully predicted an interaction with several cancer-related miRNAs. Finally, a further KEGG analysis showed that PPAR, Axon guidance, Wnt and Cell cycle signaling pathway were key putative pathways in the process of LSCC. hsa_circ:chr20:31876585-31,897,648 was found to be able to differentiate most of LSCC from the matching normal tissues. This observational study demonstrated dysregulation of circRNA in LSCC, which may have an impact on development of potential biomarkers in this disease. Validation of down-regulation of hsa_circ:chr20:31876585-31,897,648 in LSCC compared to each adjunctive tissue by Q-RT-PCR, indicating that hsa_circ:chr20:31876585-31,897,648 may be a novel promising tumor suppresser in LSCC.

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Circular RNA expression profiling identifies specific circular RNAs in tongue squamous cell carcinoma.

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Circular RNA expression profiling identifies specific circular RNAs in tongue squamous cell carcinoma.

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