The Differential Expression Of Circular RNA In Peripheral Blood Of Patients With Primary Liver Cancer In The Perioperation Of Liver Transplantation

The liver is an extremely important metabolic organ in the body,its the dysfunction of function will affect the normal metabolism of body and changes of internal environment.Primary liver cancer?PLC?is a class of malignant tumors that occur in the liver,which is generally developed by chronic liver disease or cirrhosis caused by chemical carcinogens or environmental factors.But the pathogenesis of PLC has not been elucidated at present,which has a certain degree of limitations for pathological diagnosis and treatment.Liver transplantation of allogeneic is the most thorough and effective method for the treatment of PLC.However,it 's a problem that how to raise the utilization rate of the donor liver and the survival rate of the recipients in the medical field.Therefore,finding new biological targets for early diagnosis and treatment assessment of PLC has became an urgent problem.In recent years,more and more researchers have used technology to explore the relationship between various biological small molecules and diseases with the development of gene chip technology of molecular biology.The circular RNA?circRNA?is a hot topic in the study of the molecular mechanism of major diseases.The circRNA is a kind of endogenous non-coding RNA with good conservatism,high specificity and capability of regulating the target gene.The expression profiles of circRNAs are carried out by using the circRNA microarray technique.Then,the differentially expressed circRNAs are screened out as a disease-related biomarkers,and are researched function.It's meaning that we can well understand the molecular mechanisms of disease,and improve the level of prevention and diagnosis of related diseases.Purpose:We used the circRNA microarray chip to obtain the expression profiles of circRNAs in peripheral blood of liver transplant patients with PLC during perioperative and healthy controls.The circRNA-related specific expression of PLC and early recovery of liver function after transplantation were screened by comparing the differences in the expression of circRNA between the patients with before and after liver transplantation and the healthy controls.It will provide a new information for early diagnosis,prognosis and treatment of PLC.Method:We collected the clinical data and peripheral blood samples from 4 patients who were diagnosed with PLC,underwent liver transplantation and had postoperative recovery between 12/01/2014 and 08/31/2015 at the Guilin 181st Hospital.The peripheral blood samples of patients were divided into preoperative group,and the patients in the postoperative group?1-day postoperative patients,3-day postoperative patients,and 7-day postoperative patients?.We also collected 4 peripheral blood samples of healthy individuals which matched to PLC age,gender and race matched as normal controls from the Physical Center of the Guilin 181st Hospital.The total RNAs were extracted by Trizol.The RNAs degradation and the concentration of RNAs were detected by using NanoDrop ND-1000.Then,circRNA microarray was used to analyze the expression of circRNAs in patients with liver transplantation during perioperative period and healthy controls.We compared the differential expression of circRNAs between the preoperative group,the postoperative group of liver transplantation and the healthy control group,and screened out the circRNAs which were correlated with the special expression of PLC and the early recovery of liver function after transplantation.Finally,RT-PCR was used to verify the accuracy of circRNA microarray analysis,and the miRNA binding site prediction software was used to predict and analyze the circRNAs related to miRNAs adsorption,and the function of circRNAs.Results:?1?In study of comparing preoperative with postoperative of transplantation,the 120 circRNAs were up-regulated expression in post transplantation?fold change?2.0?,and the 182 circRNAs were down-regulated expression in post transplantation?fold change?-2.0?.?2?The study of pre&post transplantation and healthy control group.The preoperative group comparing with the healthy control group was up-regulated?fold change?2.0?and the postoperative group comparing with the preoperative group was down-regulated?fold change?-2.0?,which screened 165 circRNAs.The preoperative group comparing with the healthy control group was down-regulated?fold change?-2.0?and the postoperative group comparing with the preoperative group was up-regulated?fold change?2.0?,which screened 80 circRNAs.?3?The 165 circRNAs were up-regulated in the presence of tumors in the body,which were down-regulated after liver resection and liver transplantation.They may be PLC-related biomarkers.The miRNA binding site prediction software showed that hsa_circRNA₁02347 had binding site with miR-203.The miR-203 was reported that it had specific expression in liver cancer.The 80 circRNAs were down-regulated in the presence of tumors in the body,which were up-regulated after liver resection and liver transplantation.They may be related to the recovery of liver function after liver transplantation.The miRNA binding site prediction software showed that the hsa_circRNA₄00031 had binding site with miR-483-5p.The miR-483-5p was reported that it was associated with the occurrence and development of liver cancer??4?The results of RT-PCR showed that the expression of hsa_circRNA₄00031,hsa_circRNA₁02032,hsa_circRNA₁03096 and hsa_circRNA₁02347 were consistent with circRNA microarray analysis.The expression levels of hsa_circRNA₁02032,hsa_circRNA₁03096 and hsa_circRNA₁02347 before liver transplantation were significantly higher than the healthy control group?p<0.01?,and showed a significant downward trend at 1 day,3 days,and 7 days after liver transplantation.Conclusion:?1?There were a large number of differentially expressed circRNA before and after liver resection and liver transplantation in PLC patients.They interact with miRNA,which may be closely related to the pathological process of PLC and the therapeutic effect of transplantation.?2?The hsa_circRNA₄00031,hsa_circRNA₁02032,hsa_circRNA₁03096 and hsa_circRNA₁02347 may be biomarkers for early diagnosis of PLC and prognosis after liver transplantation.