Expression And Clinical Significance Of CircRNA In Cervical Cancer And Paracancerous Tissue Based On Gene Chip

BackgroundAfter 100 years of extensive radical operation of cervical cancer,the technology has been standardized and mature.The curative effect of early stage cervical cancer patients is clear and the five year survival rate can be as high as 80%.However,the recurrence and metastasis of the cervical cancer is still a serious threat to the life of the patients.The imaging examination results in the recurrence and metastasis of the tumor,which can be shown after the formation of the tumor.It is found that the time is late,the accuracy is low,the early prediction can not be made,and the curative effect of the recurrence and chemotherapy is poor.For patients with cervical cancer after systematic and standardized treatment,there is no stable and effective method to predict the timing of postoperative recurrence and metastasis.The underlying reason is that we are not entirely clear about the gene expression and molecular regulation mechanism of cervical cancer.Therefore,it is particularly necessary to search for specific and sensitive molecular markers to objectively and accurately assess and monitor the prognosis of cervical cancer.In recent years,RNA(circular RNA,circRNA)with the closed loop structure in eukaryotic cells has been found in cancer with the significantly different expression,such as colorectal cancerliver cancer,esophageal cancer and so on,which is closely related to the occurrence and development of tumor.It may become an ideal molecular marker and is used for the diagnosis and prediction of tumor.Up to now,there are few reports about circRNA and cervical cancer.Whether circRNA exists in cervical cancer is not much known as biological function,and it is more difficult to predict the recurrence and metastasis of cervical cancer as early as possible.ObjectiveThe purpose of this study is to select the differential expression of circRNA in cervical cancer tissue by using chip technology,verifying by RT-qPCR,analyzing the clinical significance of differentially expressed circRNA with cervical cancer,laying the foundation for accurate prediction of cervical cancer.Methods1.Select,verify and analyze the differential expression of circRNA in cervical cancer tissues and adjacent tissues10 patients with cervical cancer were randomly selected to undergo extensive radical resection of cervical cancer.The differential expression profiles of cervical cancer and adjacent tissues were screened by high throughput circRNA chip technology,and the differential expression of circRNA was verified by RT-qPCR.The Gene Ontology(GO)analysis of the target genes and related genes corresponding to the significant differential expression of circRNA was analyzed,and the biological process of circRNA in cervical cancer was analyzed,and the relationship between the differential expression of circRNAs and the development of cervical cancer was explored,which could be helpful to understand the detailed pathogenesis of cervical cancer.It can provide guidance for the prognosis and evaluation of cervical cancer.2.Analyze the correlation between differential expression of circRNA and routine prognostic indicatorsBased on last part of the study,circRNA was selected by RT-qPCR technology to detect the relative expression level in the other 28 cervical cancer tissues,and the relative expression level was analyzed by statistical methods with the HPV infection,tumor size,histological type,cell differentiation degree,FIGO staging,and recurrence and metastasis of cervical cancer to determine whether circRNA can be used as a molecular marker for evaluating the prognosis of cervical cancer.Results1.Differential expression of circRNA expression in cervical cancer and its matched normal tissuesThe supervised hierarchical cluster analysis showed that the differential expression of circRNA could be correctly distinguished between the tumor tissue and the para cancerous tissue of the cervical cancer.An average of 25749 circRNA between cervical cancer and normal tissue(13304~41830)can be differentially expressed(more than 2 fold change),including 12806 circRNA(7366~20036)whose expression level is up more than 2 times,12943 circRNA(6467~21794)whose expression level is decreased more than 2 times.2 Verification the differential circRNA by real-time quantitative PCR(RT-qPCR)Hsa-circ-0013058,hsa-circ-0026919,hsa-circ-0022812,hsa-circ-0018917,hsa-circ-0079474,hsa-circ-0069272 were randomly selected,which were differentially expressed in cervical cancer,using real-time quantitative PCR(RT-qPCR)technology to verify the expression level of circRNA.The results showed that 78.6%(11 / 14)circRNAs were consistent with those by microarray.3 Bioinformatics analysisUsing GO analysis to analyzing the relationship between the differential expression of circRNA,mRNAs and miRNAs in cervical cancer,knowing the signal pathway involved in the analysis of circRNA.It was showed that the differential encoding genes and persistent gene were mainly involved in cells encoding process,such as cell cycle,cell migration and cell movement,finding circRNA was involved in the occurrence and development of cervical cancer with the interaction on relevant m RNA and miRNA.4 The relationship between the differential expression of circRNA and clinical indicators in cervical cancerThe high expression of hsa-circ-0022812,low expression of hsa-circ-009272 in cervical cancer were chosen,analyzing their correlation with HPV infection,tumor size,celldifferentiation,histological type and postoperative recurrence and metastasis.The results showed that the expression of hsa-circ-0022812 was closely connected with HPV infection,cell differentiation,tumor size,and postoperative recurrence and metastasis(P < 0.05);the expression of hsa-circ-009272 was closely connected with HPV infection,cell differentiation,histological type and postoperative recurrence and metastasis(P < 0.05);their differential expression were closely connected with HPV infection,cell differentiation and postoperative recurrence and metastasis in cervical carcinoma,suggesting that circRNA can be used to evaluate the efficacy and prognosis of cervical cancer as a molecular markers.ConclusionsCompared with the adjacent tissue,the expression of circRNA in cervical cancer were significantly changed,which was verified by RT-qPCR.GO analysis showed that circRNA,which was differentially expressed,was involved in the expression of oncogenes such as cell cycle,cell migration and cell motility,and it was related to the interaction of target genes,m RNAs and miRNA,which directly or indirectly participated in the carcinogenesis process of cervical cancer.In addition,there was significant correlation between the differential expression of circRNA and clinical evaluating index,such as HPV infection,cell differentiation,recurrence and metastasis.It is suggested that circRNA can be used to assess the curative effect,evaluate the prognosis and determine the potential therapeutic target as a molecular marker.