| Cholangiocarcinoma refers to the malignant tumor originating from the epithelial cells of the biliary system.According to its anatomical location,it can be divided into intrahepatic cholangiocarcinoma and extrahepatic cholangiocarcinoma.The incidence of cholangiocarcinoma accounts for about 3% of all gastrointestinal malignancies,and its incidence increases year by year worldwide.The malignant degree of cholangiocarcinoma is high,the condition is concealed,and the diagnosis is difficult.The patient’s condition often develops into the middle or late stage of cancer at the time of diagnosis.At present,the most effective treatment for cholangiocarcinoma is mainly surgical resection.However,due to the biological characteristics of cholangiocarcinoma,the best time for surgery has often been lost,and only one-third of patients can be treated with surgery.For patients with cholangiocarcinoma who cannot undergo surgery or postoperative recurrence,combined chemotherapy is usually used for treatment.However,cholangiocarcinoma has a high ability to promote connective tissue proliferation,and its tumor microenvironment is extremely rich.These characteristics determine its resistance to chemotherapy drugs and limit the therapeutic effect of chemotherapy.More and more studies have found that autophagy plays an important role in the development,metastasis,recurrence and drug resistance of a variety of tumors,including cholangiocarcinoma.There have been many studies on autophagy in cholangiocarcinoma,but there are still many controversies about the mechanism of autophagy in cholangiocarcinoma.According to the different entry points,the results still remain undifferent.Objective: To investigate the effect of Irisin on cholangiocarcinoma(Hucct-1):observe the activity of Hucct-1 cells and the level of autophagy,and further detect the changes of autophagy-related signaling pathways,and to explore the possible mechanism of Irisin on cholangiocarcinoma.Methods: A total of 37 surgically resected and pathologically confirmed cholangiocarcinoma specimens from the Department of Hepatobiliary Surgery,Qingdao Municipal Hospital fromSeptember 2016 to September 2018 were selected.The immunohistochemical method was used to detect the level of Irisin in cholangiocarcinoma specimens;CCK-8 assay was used to detect the effect of Irisin on the proliferation of cholangiocarcinoma in cultured cells(Hucct-1);scratch test and Transwell assay were used to detect the migration and invasive ability of Hucct-1;the change of autophagy level was detected by monodansylcadaverin(MDC)staining method;the autophagy-related protein LC3 B and autophagy-related channel PI3K/ Akt/mTOR were detected by Western-blot.Results: Immunohistochemical results showed that patients with low Irisin expression accounted for 51.4%(19/37),moderate expression accounted for35.1%(13/37),and high expression accounted for 13.5%(5/37),indicating Irisin has different degrees of expression in cholangiocarcinoma,and it is important to investigate the effect of Irisin on cholangiocarcinoma.The results of CCK-8 showed that Irisin inhibited the proliferation of Hucct-1,and the effect was related to the dose of Irisin.Scratch test showed that the cell healing rate of cells with Irisin(33±3)% was significantly lower than control group(45±4)%,and the results were statistically significant(p<0.05).The Transwell experiment showed that the number of invasive cells in the Irisin treatment group was(82±5),which was significantly lower than that in the control group(146±8).The results were statistically significant(p<0.05).MDC staining showed that the cells treated with Irisin had higher autophagy activity.Western-blot results also showed that the level of autophagy-related protein LC3 B increased after Irisin treatment,and the levels of Akt and mTOR proteins in the PI3K/Akt/mTOR pathway decreased,indicating that Irisin has a negative regulatory effect on this pathway.Conclusion:Irisin can promote the proliferation,migration and invasion of Hucct-1 by promoting autophagy via PI3K/Akt/mTOR signaling pathway。... |
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