| Objective: Tongue squamous cell carcinoma(TSCC)is one of the most common malignant tumors in the oral and maxillofacial regions in China.TSCC is characterized by difficulty to cure,metastasizes easily,and poor prognosis.At present,the mechanism of TSCC development remains unclear.Recent studies have shown that circularRNA(circRNA)can play a critical role in tumorigenesis and tumor progression.However,the role of circRNA and its biological function and mechanism in the development of TSCC is far from clear.To better understand circRNA expression patterns in TSCC,in this study,circRNAs expression profiles in TSCC were analyzed and identified by high-throughput sequencing and circRNA microarray.Various bioinformatics strategies and methods were used to analyze the abnormal circRNAs expression in TSCC.The mechanism of circRNAs in TSCC was explored by bioinformatics analysis and experiments,providing a new perspective and theoretical basis for the clinical diagnosis and targeted therapy of TSCC,facilitating the development of new prognostic biomarkers and therapeutic targets for TSCC.Methods:(1)High-throughput sequencing and circRNA microarray were used to explore circRNAs in TSCC and their adjacent normal tissues.The expression of circRNAs in TSCC was analyzed by heat map,volcano map,scatter map,and other strategies and the results were validated in TSCC samples by qRT-PCR.(2)The target circRNA was selected by bioinformatics analysis,including GO,KEGG and Veen Diagram,GEPIA2,and UALCAN databases.(3)qRT-PCR was used to validated and analyzed the expression level of hsa＿circ＿0125480 in TSCC tissues and cell lines.(4)The human TSCC cell lines were transfected with overexpression plasmid of hsa＿circ＿0125480.The effects of hsa＿circ＿0125480 overexpression on proliferation,invasion,and migration in TSCC cells were evaluated by CCK-8,clone formation,wound healing,and transwell invasion assay.(5)The circRNA/miRNA/mRNA network was established,and the potential biological role of hsa＿circ＿0125480/miRNA/mRNA network in TSCC was revealed by using protein-protein interaction map,GO enrichment,and KEGG pathway analysis.(6)The role of hsa＿circ＿0125480/mi R-766-3p/NR3C2 axis in TSCC was explored by bioinformatics prediction and assessed by the co-expression level of hsa＿circ＿0125480,mi R-766-3p and NR3C2 in TSCC samples and TSCC cell lines.Results:(1)Through high-throughput sequencing,277 differentially expressed circRNAs were found,of which 69 were significantly up-regulated and 208 were significantly downregulated in TSCC tissues;trough circRNA microarray detection,124 circRNAs differentially expressed were found,of which 54 were significantly up-regulated in TSCC tissues and 70 were significantly down-regulated in TSCC tissues.(2)Multiple abnormally expressed circRNAs in TSCC are involved in tumor-related signaling pathways.(3)hsa＿circ＿0125480 was significantly low expressed in TSCC tissues and cell lines and was a potential biological marker of TSCC.(4)Overexpression of hsa＿circ＿0125480 in TSCC cells inhibited the malignant behavior of TSCC cells.(5)hsa＿circ＿0125480 can interact with multiple miRNAs and mRNAs and participate in multiple tumor-related pathways.(6)The expression level of hsa＿circ＿0125480,NR3C2,and the related target mi R-766-3p in TSCC specimens and cell lines were correlated,indicating the role of hsa＿circ＿0125480/mi R-766-3p/NR3C2 axis in TSCC.Conclusion: In this study,the expression profile of circRNAs in TSCC was analyzed by high-throughput sequencing and microarray.The expression levels of differentially expressed circRNAs were validated in TSCC samples.The validation results in TSCC patients showed that the expression tendency of differentially expressed circRNAs was consistent with the results of high-throughput sequencing and microarray,and hsa＿circ＿0125480 was significantly down-expressed in TSCC tissues.According to bioinformatics analysis results,we selected hsa＿circ＿0125480 for further study and found that overexpression of hsa＿circ＿0125480 in TSCC cells inhibited the malignant behavior of CAL27 and SCC9.Combined with competitive endogenousRNA theory,the result of bioinformatics analysis,and molecular experiments,we found that hsa＿circ＿0125480/mi R-766-3p/NR3C2 signaling axis may be involved in regulating the biological behavior of TSCC.Our study suggested that hsa＿circ＿0125480 may be a critical prognostic marker and a potential therapeutic target for TSCC. |
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