TGF-?1/Smad Signaling Pathway Mediated Epithelia-mesenchymal Transitian In The Fibrosis Of Hashimoto's Thyroiditis

ObjectiveTo investigate the mechanism of TGF-?1/Smad signaling pathway mediated epithelialmesenchymal transition(EMT)in Hashimoto's thyroiditis fibrotic process;On the basis of organization and cytology vitro experiments,this paper explores EMT-related markers and TGF-?1/Smad signaling pathway proteins expression and function in the development and progression of Hashimoto's thyroiditis,and analyze the relationship with it's clinicopathological features.We further aimed to study the function of TGF-?1/Smad signaling pathway in EMT of Hashimoto's thyroiditis,and lay the foundation for transformation mechanism in HT,and to provide a theoretical basis for the blocking and even reversal of Hashimoto's thyroiditis fibrosis treatment.Methods1.47 cases of Hashimoto's thyroiditis surgical specimens were detected the expression of EMT related protein and TGF-?1/Smad signaling pathway proteins by immunohistochemical method using tissue microarray.Then,the correlation between the proteins and its contact with Epithelial Mesenchymal Transition in the follicular epithelial cells was analyzed.2.Different concentrations of rh-TGF-?1 cytokines and the inhibitors of TGF-?1/Smad signaling pathway-rh-BMP7 dealed with normal thyroid epithelial cells-Nthyori-31,using an inverted microscope observed the changes in cell morphology,Real-Time PCR and Western Blotting examined the expression of TGF-?1/Smad signaling pathway markers and EMT markers in the cell line Nthyori-31 induced by rh-TGF-?1 and rh-BMP7 in the different condition.Results1.Immunohistochemical detection showed that the positive expression of TGF-?1 in the Fibrous variant of Hashimoto's thyroiditis(FVHT)had a significant difference(?2 =17.92,p <0.01)with the the typical Hashimoto's thyroiditis.The positive expression rate of TGF-?1 in FVHT was significantly higher than the typical HT.The expression of Smad2/3and TGF-?1 is consistent(Kappa=0.783,p<0.05).The expression of E-cadherin was significantly reduced and the expression of Vimentin was significantly increased and the correlation between them was negative(r=0.549,p<0.05).The reduced expression of E-cadherin had not correlation with the positive expression of TGF-?1(r=0.024,p>0.05)and the increased expression of Vimentin has correlation with the positive expression of TGF-?1(r=0.282,p <0.05)in Hashimoto's thyroiditis.2.The Nthyori-31 cells dealed with different concentrations of rh-TGF-?1 had morphological changes of epithelial cells to mesenchymal cells,the fluorescence quantitative PCR detection showed that the RNA levels of Vimentin,?-SMA,Smad2,Smad3 and Smad4 were significantly up-regulated,the RNA levels of E-cadherin and Smad7 were significantly down-regulated.The rh-TGF-?1 and rh-BMP7 treated Nthyo-ri-31 cells at the same time,the fluorescence quantitative PCR and Western Blot detection showed that BMP7 can inhibit the up-regulated expression of Vimentin,?-SMA,Smad2,Smad3 and Smad4 and the down-regulated expression of E-cadherin and Smad7.Conclusions1.The epithelial-mesenchymal transition(EMT)phenomenon is prevalent in Hashimoto's thyroiditis,but we still can not believe that this phenomenon has correlation with the positive expression of TGF-?1.Compared with typical HT,TGF-?1 and Smad2/3 all have significance higher rate of positive expression,all the above showed that TGF-?1/ Smad signaling pathway plays an important role of regulatory mechanism in Hashimoto's thyroiditis fibrosis.2.TGF-?1 can induce epithelial-mesenchymal transition in normal thyroid epithelial cells Nthyori-31 through TGF-?/ Smad signaling transduction pathway.