The Inhibition Effects Of Allicin On Epithelial-mesenchymal Transition Of Cholangiocarcinoma Cells

Objective:This study focused on investigating the inhibition of allicin on epithelium mesenchymal transition(EMT)of human cholangiocarcinoma cells and its related mechanism,and providing theoretical basis for the application of allicin in the treatment of cholangiocarcinoma.Methods: MTT assay were used to detect the inhibitory effect of allicin on the human cholangiocarcinoma RBE cell proliferation,and the drug concentration of allicin was determined by IC50 of 24 h.The RBE cells were cultured and divided into control group,allicin group(IC50),TGF-b1 group(10ng/m L)and allicin+ TGF-b1group(IC50+10ng/m L).Wound scratch test and Transwell invasion assay were used to performed to detect the migration and invasion ability of RBE cell after 24 h.Western blot were applied to detect expression of EMT-related proteins(E-Cadherin,N-Cadherin,Vimentin,Snail)and NF-?B signaling pathways.Results: The results showed that allicin could inhibit the proliferation of RBE cell in a dose-dependent manner,and the IC50 of 24 h was 130.7?mol/L.The Wound scratch test showed the migration rates at 24 h in allicin group and allicin+TGF-b1 group was decreased compared with that in the control group(9.25% ± 0.36% vs 28.19 %±0.66%,P<0.05)and TGF-b1 group(13.91% ± 0.75% vs 49.22 %± 0.27%,P<0.05).Transwell invasion assay experiment showed the invasion rates at 24 h in allicin group and allicin+TGF-b1 group were also decreased compared with that in the control group(6.59% ± 0.06% vs 33.48%± 0.04%,P<0.05)and TGF-b1group(9.40% ± 0.05% and 40.21 %± 0.12%,P<0.05).Western showed that compared with the control group,Snail,N-Cadherin,Vimentin,NF-kB and p-NF-kBexpression were significantly decreased in the allicin group,and E-Cadherin expression was significantly increased(P<0.05).Compared with TGF-b1 group,E-Cadherin expression was significantly up-regulated,and Snail,N-Cadherin,Vimentin,NF-kB and p-NF-kB expression were significantly down-regulated in the allicin+TGF-b1 group(P<0.05).Conclusion: These results indicate that allicin can inhibit the proliferation of cholangiocarcinoma RBE cells.Allicin inhibit the EMT of human cholangiocarcinoma cells by blocking NF-?B signaling pathway and down-regulating the expression of Snail,N-Cadherin,Vimentin and up-regulating the expression of E-Cadherin.Allicin counld also inhibit the EMT induced by TGF-?1 on the human cholangiocarcinoma cell,which may have potential value to be the drug candidate for the treatment of human cholangiocarcinoma in future.