Expression Of MicroRNA-375 And Nuclear Factor-κB In Thyroid Papillary Carcinoma And Their Significance

BackgroundThyroid carcinoma(TC),which has become the first occurrence rate of endocrine malignant tumor and the seventh occurrence rate of all tumors in China,is important nowadays.Papillary Thyroid Carcinoma(PTC)also matters because of it’s high ratio.The 5-year survival rate for PTC patients is over 90%,most patients achieve good clinical outcomes after receiving standardized surgery and combined treatment.However,5%-10%PTC patients is more aggressive due to recurrence and distant metastasis occurring in the early stage.It then progresses to another kind of malignant disease which would generate adverse effects on patients,called radioactive iodine refractory TC.So people affirmed the significance of PTC research.MicroRNAs(miRNAs)is found in a lot of cells with no ability to encipher.MiRNAs could affect the expression of protein through binding to Messenger.Ribonucleic Acid(mRNA).MicroRNA-375(miRNA-375),as one of the members of miRNAs,plays a role in a lot of diseases.Nuclear factor-κB(NF-κB)gets a number of capacities.For example,in the period of growth of body,it could change the process of embryonic development by altering multiplication and differentiation of cells.And when body get in the stress and ill progression,it could improve the react of defence system.However,if the balance were broken,the transformation of tumor could also be induce.In recent years,there are many researches proved that the tumor progression would change with the unusual expression of miRNA-375.NF-κB get the same ability.Some scholars have indicated that there is a regulatory relationship between miRNA-375 and NF-κB pathways.However,there are little researches concerning the miRNA-375 and NF-κB’s role in TC.The goal of us is to test miRNA-375 and P65 in PTC,and their connection.The trait of PTC involving the two was also studied.Purpose1.The miRNA-375 in PTC tissues and paracancer tissues was tested.The link between miRNA-375 and clinical characteristic of PTC was then explored.2.The P65 in PTC and paracancer tissues was tested,the P65 was tested in TPC-1 cells.3.To analyze the relationship between miRNA-375 and NF-κB signaling pathways in thyroid papillary carcinoma.MethodFrom January 2018 to December 2018,60 patients undergoing TC surgery in the Thyroid Surgery Department of the First Affiliated Hospital of Zhengzhou University were selected.The agreements were get before operation.Everyone signed informed consent of sampling from the biobank.We gathered PTC tissues and normal tissue.The RT-qPCR was used to measure miRNA-375 in all tissues.WB was used to explore the P65 in 22 pairs of tissues.We constructed the TPC-1 cell model transfected with lentivirus,and then we established the blank control.We divided four groups:BC,miR-con,miR-375 and miR-375 inhibitor.Dual luciferase report analysis system was used to detect RELA activity.WB was used to find the protein levels of several groups of P65.MTT assay was used to judge the proliferation activity of different groups.We applied t test to mouse out the imparity of miRNA-375 and P65 in PTC and tissues.We applied Mann-whitney U and t-test seek the link between miRNA-375 and the clinical features of PTC.We applied ROC curve to find the diagnosibility of miRAN-375 for PTC.We applied Pearson method to exam the link between miRNA-375 and P65.In the cell test,the t test was used for comparison between the two groups.ResultWe can notice from RT-qPCR that the PTC tissues got a lower miRNA-375 than adjacent normal tissues(P<0.01).When a patient has a lymph metastasis or local tumor invasion,the level of miRNA-375 of PTC tissues become lower.(P=0.038,0.08).WB revealed that the P65 in PTC tissues was higher than normal tissues(P<0.01).When we use 0.8842 of miRNA-375 for finding PTC,the sensitivity and specificity were 86.7%and 71.7%,respectively,and the Jorden index was 58.3%.MiRNA-375 level was correlated with NF-κB in PTC(rs=-0.791,P<0.01).In TPC-1 cells,the fluorescence expression intensity of RELA decreased when miRNA-375 was overexpressed,and increased when miR-375 was inhibited(P<0.05).The P65 protein showed the same trend(P<0.05).The results of MTT assay showed that the cell growth rate was decreased when miRNA-375 was overexpressed in TPC-1 cells,and when miRNA-375 was inhibited,the result opposite(P<0.05).ConclusionmiRNA-375 expression was reduced in PTC,and its level was related to progression of PTC.The miRNA-375 in the tissue has certain sense for the comfirmation of PTC.NF-κB pathway is activated in PTC and TPC-1 cell.There is a negative relationship between the two,and maybe a regulatory relationship.The miRNA-375 could inhibit the proliferation of TPC-1 cells in vitro.