Effict Of Cyclopamine Inhabited Hedgehog Signaling Pathway On The Rewersion Of Epithelial-Mesenchymal Transition In Esophageal Cancer Cells And Its Mechanism

Objective:To explore the effect of cyclopamine on epithelial-mesenchymal transition in human esophageal cancer EC 109 cells and the possible mechanism.Methods:EC109 cells were cultured in vitro. After adding cyclopamine 48h, the expression of Gli1 was tested by real-time PCR (Fluorescent quantitation polymerase chain reaction) and morphological change of cells was observed by inverted microscope.The transwell chamber assay was used to examine the invasive and metastatic ability of EC 109 cells treated with cyclopamine for 48h.The ablity of vascularization was observed by the method of vascilogenic mimicry and the ablity of adhension was detected by adhension experiment. Real-time PCR and Western blot were used to detect the expressions of such EMT markers as E-cadherin, β-catenin and vimentin and such transcription factors as Snail, Twist 1.Results:The inhibition of hedgehog signaling significantly reduced the mRNA expression of Gli1 (41.819±20.150)%,and the invasion[(24.800±2.588)vs.(55.400±4.879)], migration capacities [(23.200±1.924)vs.(65.400±4.775)] of EC109 cells in vitro were decreased(t=-12.390,P= 0.000; t=-18.331,P=0.000).And homogeneous cells intercellular adhesion was increased(F=9.327, P=0.009),while heterogeneous cells intercellular adhesion was decreased (F=20.459,P=0.000).And angiogenesis capacity [(2.780±0.424)vs (5.080±0.634)]was decreased(t=-16.919,P=0.000).The expression of E-cadherin increased [(0.3877±0.5648)vs.(0.2275±0.5823)] significantly (t=3.421,P=0.027),while Vimentin and β-catenin in the cyclopamine treatment group [(0.5877±0.1091)vs.(0.5071±0.0509) (0.9983±0.1284) vs.(0.7556±0.0379)]were significantly down-regulated (t=4.221,P=0.013; t=6.781,P= 0.002). Compared to the control group, the expressions of Snail were significantly reduced in the cyclopamine treatment group [(0.7556±0.0379) vs. (0.4013±0.0205)] (t=-6.455,P=0.023).Also,the mRNA expressions of Twistl significantly reduced (74.987±9.031)%.Conclusion:The inhibition of hedgehog signaling pathway in vitro can effectually suppress the process of EMT,even reversed this process. The mechanism is partially correlated with the down-regulations of Snail and Twist1.