The Study On The Diagnostic Value Of Serum Free Circular RNAs In Detecting Early Nasopharyngeal Carcinoma

Objective:Early diagnosis of nasopharyngeal carcinoma is crucial for prognosis.CircRNA has been reported as a biomarker of cancer,and many studies have reported the mechanism of circRNA on the occurrence and development of nasopharyngeal carcinoma.However,whether there is a specific expression profile in the serum of NPC patients,the diagnostic value of circRNA specifically expressed in NPC patients,and the mechanism of the progression of NPC are not clear.This study aims to explore whether there is a specific expression profile in the serum of patients with early nasopharyngeal cancer,and then to explore the diagnostic efficacy of differentially expressed circRNA in nasopharyngeal cancer and its mechanism of action in the occurrence and development of nasopharyngeal cancer,so as to find a breakthrough for the pathogenesis of nasopharyngeal cancer.Methods:1.CircRNA microarray technique was used to detect the expression profile of circRNA in 16 serum samples,and the specific expression profile in nasopharyngeal carcinoma was further analyzed.The top five circRNAs with up-regulated and down-regulated expression were selected and verified by qRT-PCR in 16 samples to verify the reliability of the chip results.2.qRT-PCR was also used to detect the expression of circRNA candidate in serum of other patients with nasopharyngeal carcinoma,patients with rhinitis nasal polyp and healthy controls,so as to evaluate its diagnostic value for early nasopharyngeal carcinoma.3.Bioinformatics techniques were used to analyze the possible role of parent genes of differentially expressed circRNAs in the pathogenesis and development of nasopharyngeal carcinoma.Meanwhile,TargetScan and miRanda databases were used to predict the miRNA and mRNA bound to circRNAs with top 10 up-regulated and down-regulated circRNAs in serum.Results:1.CircRNA microarray technology found differentially expressed circRNAs in the serum of patients with early nasopharyngeal carcinoma,including 177 significantly up-regulated circRNAs and 179 significantly down-regulated circRNAs,suggesting that circRNA has a variety of unclear functions in the occurrence and development of nasopharyngeal carcinoma.Through screening,we obtained three candidate circRNAs with differential expression in early nasopharyngeal carcinoma,and verified by PCR technology and product sequence,indicating that the candidate circRNA does have differential expression in early nasopharyngeal carcinoma serum.2.Compared with healthy controls,the expression of hsa＿circ＿0006220 was significantly up-regulated in nasopharyngeal carcinoma patients,and the area under the curve of serum hsa＿circ＿0006220 against early NPC was 0.8260（95%CI 0.6922-0.9598）.The expression of hsa＿circ＿0006220 was also significantly up-regulated in nasopharyngeal carcinoma patients compared with the rhinitis nasal polyp group,and the area under the curve of serum hsa＿circ＿0006220 against early NPC was 0.7917（95%CI 0.64610.9373）.The same circRNAs candidate hsa＿circ＿0001666 was significantly upregulated in nasopharyngeal carcinoma patients compared with healthy controls.The area under curve of serum hsa＿circ＿0001666 against early NPC was 0.8933（95%CI 0.9135-1.000）.The expression of hsa＿circ＿0001666 was also significantly up-regulated in nasopharyngeal carcinoma patients compared with the rhinitis nasal polyp group,and the area under the curve of serum hsa＿circ＿0001666 against early NPC was 0.8200（95%CI 0.6797～0.9603）.Hsa_circ₀006220 and hsa＿circ＿0001666 have good diagnostic value for NPC in early stage,and the important findings of this study reveal the role of peripheral blood free circRNAs in NPC diagnosis,which will be conducive to the discovery of new molecular diagnostic markers for NPC with potential clinical application.3.Bioinformatics techniques were used to analyze the potential role of circRNAs parental genes differentially expressed in serum in the occurrence and development of nasopharyngeal carcinoma,and predict the binding of 5 miRNAs and their downstream mRNAs.Conclusions:There is specific circRNAs expression profile in the serum of early nasopharyngeal carcinoma patients,and the two up-regulated circRNAs selected from the differentially expressed circRNAs expression profile in the serum of early nasopharyngeal carcinoma patients have moderate diagnostic value.However,both up-regulated and down-regulated circRNAs in serum of early NPC are involved in the occurrence and development of NPC,especially the patch adhesion signaling pathway.We believe that circRNAs not only have diagnostic value for NPC,but also may play an oncogene role in the occurrence and development of NPC.