Effect And Molecular Mechanisms Of IL-23/miRNA-25/SOCS4 Signaling Pathway In Papillary Thyroid Carcinoma Metastasis

Background:Thyroid cancer,whose morbidity has an increasing trend year by year,is a common head and neck neoplasm.The survival rate of thyroid cancer patients who had undergone surgical treatment and postoperative comprehensive treat is relatively high,but is still quite higher than any other pancreatic endocrine tumors.Furthermore,the therapeutic effect of thyroid cancer patients with metastasis is far more than those without.It is very meaningful to explore the molecular mechanism of metastasis of thyroid cancer for the advance of overall therapeutic effect.In recent years,the important role of interleukin in the metastasis of tumors has obtained more and more concern from researchers.It has been reported that interleukin-23(IL-23)is up-regulated in many types of tumor tissues and related closely to tumor metastasis.However,the role of IL-23 in the occurrence and development of thyroid cancer is still not very clear.Hence,the investigation about the role of IL-23 in the occurrence and development of thyroid cancer has practical significance for the prevention and treatment of thyroid cancer.Whether IL-23 is related to the occurrence of thyroid cancer?Whether IL-23 could affect the metastasis of thyroid cancer?What is the molecular mechanism about it?All of them are need to be answered in the present study.methods:1)First,we detected the expression of IL-23 protein and mRNA in the tumor tissue and normal tissue of thyroid.2)In vitro,we researched the change of proliferation,migration and invasion ability of thyroid cancer cell line K1 after being dealt with exogenous IL-23.3)After that,the change of the mRNA and protein expression of SOCS4 in K1 cell was detected through real-time PCR and western blot.4)Using biology engineering technology,we overexpressed and inhibited respectively SOCS4 gene expression in thyroid cancer cell line K1,and detected the change of migration and invasion ability in vitro.5)After the overexpression and inhibition of SOCS4 gene in thyroid cancer cell line K1,the cell migration and invasion ability was detected again after being dealt with exogenous IL-23.It was to clarify whether the regulation of thyroid cancer cells migration and invasion ability depends on SOCS4 gene expression or not.6)By using bioinformatics method we analyzed the particular microRNA of SOCS4 3’-UTR area and through luciferase reporter system it was verified whether microRNA is specifically bound with SOCS4 3’-UTR area.After being dealt with exogenous IL-23,the change of the particular microRNA in thyroid cancer cells was detected.The comprehensive analysis about the important role of particular microRNA in the regulation of IL-23 on SOCS4 had been done.7)Using particular microRNA mimics and inhibitor to deal with thyroid cancer cells,the change of SOCS4 gene expression and migration and invasion ability of thyroid cancer cells in vitro was detected.8)Using real-time PCR,the expression of IL-23,particular microRNA and SOCS4 mRNA was detected and the correlation among them was analyzed to verify their interaction.Results:1)The result of immunohistochemistry showed that IL-23 protein expression in normal thyroid tissue was negative or weak positive,in thyroid cancer tissue positive or strong positive(18/20)(p<0.01);2)After being dealt with exogenous IL-23,the proliferation of thyroid cancer cell line K1 was not changed obviously,however,migration and invasion ability depending on time and concentration in vitro was enhanced significantly(p<0.05);3)After being dealt with exogenous IL-23,the expression of SOCS4 mRNA and protein in thyroid cancer cell K1 was decreased significantly(p<0.05);4)After the targeted inhibition of SOCS4 gene,migration and invasion ability in vitro of thyroid cancer cell K1 was enhanced significantly.When SOCS4 gene was inhibited,migration and invasion ability in vitro of thyroid cancer cell K1 was decreased significantly.It showed that SOCS4 gene expression was closely related to the migration and invasion ability in vitro of thyroid cancer cells.5)The overexpression of SOCS4 gene could significantly suppress the increasing of migration and invasion ability in vitro of thyroid cancer cell K1 after being dealt with exogenous IL-23.The targeted inhibition of SOCS4 gene could reinforce the increasing of migration and invasion ability in vitro of thyroid cancer cell K1 after being dealt with exogenous IL-23.6)Through 3 websites about miRNA and target gene prediction,we analyzed 14 microRNAs that may have potential binding sites with SOCS4 mRNA 3’-UTR area.There were 6 that can decrease the fluorescence value of SOCS4 over 50%.After being dealt with exogenous IL-23,the result of detection revealed that it was may be miRNA-25 that can bind with SOCS4 mRNA 3’-UTR area.7)After being dealt with particular microRNA mimics and inhibitor of miRNA-25,the SOCS4 gene expression in thyroid cancer cell K1 had changed significantly,meanwhile,migration and invasion ability of thyroid cancer cells in vitro was also changed significantly.8)The result of real-time PCR showed that IL-23 mRNA had positive correlation with the expression of miRNA-25 and negative with SOCS4 mRNA,and that miRNA-25 had negative correlation with the expression of SOCS4 mRNA.Conclusions:1)The expression level of IL-23 in tumor tissues is significantly higher than in normal tissues.2)Exogenous IL-23 can promote the migration and invasion ability of thyroid cancer cells.3)IL-23 can inhibit the expression of SOCS4 gene through inducing the up-regulation of miRNA-25,and then promote the migration and invasion ability of thyroid cancer cells.