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Celastrus orbiculaus is one of the important folk medicinal plants used in the anti-inflammatory and analgesic treatment of various diseases.Recently, study about the ethyl acetate extract of Celastrus orbiculatus (COE) has shown that it has significant antitumor effects.However, study of the effect and mechanism of COE on suppressing the epithelial-mesenchymal transition(EMT) of human gastric adenocarcinoma cells have not been reported at home and abroad so far.This experiment mainly study that COE could inhibit EMT of gastric by regulating cell cytoskeleton rearrangement.The effect of COE on the activity of AGS cells was detected by MTT assay, and the suitable test concentrations were determined.EMT model was iduced by TGFβ1.Cell cytoskeleton staining, laser scanning confocal microscopy and electronic microscope were used to detect the morphological changes in cell morphology and microstructure of EMT cells before and after COE treatment.Invasion assay and migration assay were used to observe the effect of COE on the metastatic ability of AGS cells in vitro.The effect of COE on the expression of cofilin 1, EMT biomarkers(including E-cadherin, N-cadherin and Vimentin), matrix metalloproteinases protein(MMPS) and Cofilin 1 protein in AGS cells were examined by western blotting.The correlation between Cofilin 1 and EMT was tested by immunofluorescence technique and cytoskeleton staining combining method.The results showed that COE could significantly inhibit the process of EMT in AGS cells and was concentration dependent.In addition, COE significantly down-regulated the level of Cofilin 1 and was concentration dependent.In conclusion, these results suggested that Cofilin 1 was directly involved in the process of AGS cell EMT, and played an important role in it.COE could significantly inhibit the epithelial mesenchymal transition in AGS cells, which is likely to be achieved by inhibiting the activation of Cofilinl signalling pathway.This study may provide a basis for the development of new anticancer drugs and development of novel therapeutic strategies targeting Cofilin 1 protein.