Expression And Significance Of Annexin-A2,EMT Markers And CX-43 In Hepatolithiasis-Associated Intrahepatic Cholangiocarcinoma

Objective: To investigate the expressions of Annexin-A2,Cx-43 and the epithelial-mesenchymal transition(EMT)markers(E-cadherin,vimentin and Cytokeratin in hepatolithiasis-associated cholangiocarcinoma in an inflammatory environment and their significance.Methods:1.In 20 case of hepatolithiasis-associated cholangiocarcinoma tissues(tumor group)and 20 case of chronic inflammatory bile duct with simple hepatolithiasis tissues(inflammatory group),Immunohistochemical method(ABC method)was used to determine the expression levels of E-cadherin and vimentin in tissue samples of tumor group and inflammation group;The expression of Annexin-A2,Cytokeratin and CX-43 in tissue samples of tumor group and inflammation group was determined by Western Blotting.2.The levels of IL-6 in fasting peripheral blood samples were measured by ELISA in tumor group and inflammatory group.Result:1.Analysis of serum IL-6 concentration by one-step ELISA method:ELISA was used to determine the serum IL-6 concentration(ng / ml)in tumor group and inflammatory group.The T-test analysis revealed that the serum IL-6 concentration in the hepatolithiasis-associated cholangiocarcinoma group was significantly higher than that in the chronic inflammatory bile duct with simple hepatolithiasis group,and the difference was statistically significant(P <0.05).2.Expression of E-cadherin andvimentin detected by immunohistochemistry : Analysis of E-cadherin expression results: Under the light microscope,it can be seen that the positive expression of E-cadherin is mainly localized in the cell membrane and cytoplasm,which is yellow or brown,and its expression is significantly higher in the inflammation group than in the tumor group.The T-test analysis of the average optical density values of the tumor group and the inflammation group was statistically significant(P <0.05)?3.Expression of Annexin-A2,Cytokeratin and CX-43 by Western Blotting : Analysis of Annexin-A2 expression results : The immunoblot method showed that protein bands appeared in the tumor group and the inflammation group.The bands in the tumor group were brighter than the inflammation group.The average optical density values of the two groups were analyzed by T-test.The expression of Annexin-A2 in the tumor group was significantly higher than that in the inflammation group.The difference was statistically significant(P <0.05).Analysis of Cytokeratin and CX-43 expression results:Western blot results showed that protein bands appeared in the tumor group and the inflammation group.The bands in the inflammation group were brighter than the tumor group.The average optical density values of the two groups were analyzed by T-test.The expressions of Cytokeratin and CX-43 in the inflammation group were significantly higher than those in the tumor group.It has statistical significance(P <0.05).4.Correlation analysis : The correlation between the expression ofAnnexin-A2 and Cx-43 was analyzed by Pearson correlation analysis based on the relative average optical density value of the reference protein.The Pearson correlation analysis showed that the expression of Annexin-A2 and Cx-43 was Negative correlation(r=-0.7823,P <0.05).The correlation between the expression of Cytokeratin and Cx-43 was analyzed by Pearson correlation analysis based on the relative average optical density value of the reference protein.The Pearson correlation analysis showed that the expression of Cytokeratin and Cx-43 was positive correlation(r=0.7120,P<0.05).Conclusions:(1)EMT was found in hepatolithiasis associated cholangiocarcinoma.(2)The level of serum IL-6 in patients with hepatolithiasis associated cholangiocarcinoma was significantly increased.(3)Inhibition of CX-43 expression in hepatolithiasis associated cholangiocarcinoma may be related to EMT.(4)The expression of annexin-a2 in hepatolithiasis related cholangiocarcinoma increased,which may be related to the inhibition of expression of CX-43.(5)Under the condition of inflammatory factors,annexin-a2 / CX-43 / EMT pathway may play an important role in the development of hepatolithiasis related cholangiocarcinoma.