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ORI can inhibit pancreatic cancer cell SW1990 migration and EMT by down-regulating Wnt/β-catenin signal transduction in vitro and in vivo.Therefore, it can be potentially and effectively used in the clinical management of pancreatic cancer.
Tumour sections stained with anti-E-cadherin, anti-vimentin and anti-β-catenin antibodies revealed that ORI inhibited EMT, as well as the Wnt/β-catenin pathway in vivo.
Wnt/β-catenin signalling was inhibited by ORI using luciferase reporter assay.
ORI can decrease the β-catenin protein level not only in the nucleus, but also in the cytoplasm and the whole cell after the treatment with ORI and glycogen synthase kinase 3β (GSK3β) was increased in the ORI-treated group.
Pancreatic cancer cell lines (Aspc1, Bxpc3, Panc1, SW1990) was obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).
The cells were cultured in DMEM supplemented with 10 % FBS, 100U/ml penicillin, and 100 μg/ml streptomycin at 37 °C in a humidified atmosphere of 5 % CO cells per well).
Wound healing assay, transwell assay and spontaneous metastasis model were used to observe the migration activities.
Real-time PCR, immunofluorescence, western blot analysis and immunohistochemistry methods were used to observe the expression of genes or proteins. Real-time PCR and immuno-fluorescence analyses of epithelial-to-mesenchymal transition (EMT) markers were compared between control group and ORI group.ORI was dissolved in dimethyl sulfoxide (DMSO) to make a stock solution (10 m M) and stored at −20 °C.The DMSO concentration was kept below 0.1 % in all experiments and did not exert any detectable effect on cell growth or cell death.Less than 20 % of patients with pancreatic cancer are subjected to surgical removal, whereas other patients are typically treated with chemotherapy or chemotherapy with radiation.Pancreatic cancer is one of the few cancers in which survival has not improved substantially in 40 years.These patients have worse prognoses than those who demonstrate less evidence of EMT.