| Objective:To explore the effects of exosomes from two different sources of pancreatic cancer cells(Pa Tu8988 and Bx PC-3)on cancer associated fibroblasts.To investigate the effect of cancer associated fibroblasts on proliferation and migration of pancreatic cancer cells,and to explore the possible mechanism by which exosomes from pancreatic cancer promote the activation of cancer associated fibroblasts.Methods:1.Exosomes were extracted from the supernatant of two different pancreatic cancer cells(Pa Tu8988 and Bx PC-3).2.Exosomes extracted from pancreatic cancer were identified by electron microscopy,NTA and western blot.3.The exosomes of pancreatic cancer were co-cultured with hepatic stellate cells(LX-2),and protein expression levels related to cancer associated fibroblasts(CAFs)were determined by western blot.4.CCK8 cell proliferation assay and scratch assay were used to detect the role of exosomes from pancreatic cancer in promoting CAFs activation in cell proliferation and migration.5.Western blot was used to detect the expression levels of proteins related to CAFs activation and NF-?B pathway promoted by exosomes from pancreatic cancer.Results:1.Electron microscopy,NTA and western blot proved that exosomes were extracted from the supernatant of two kinds of pancreatic cancer cell cultures.2.The extracted exosomes were co-cultured with hepatic stellate cells(LX-2)and the expression levels of ?-SMA and FAP proteins in Cafs-related markers were increased by western blot assay.3.After co-culture with exosomes and LX-2,the proliferation and migration of cells were enhanced,and the expression of p-NF-?B protein in NF-?B pathway was increased,but the expression of NF-?B protein was not significantly changed.The expression levels of p-I?B? and I?B? decreased,especially p-I?B?.Conclusions:Exosomes from pancreatic cancer can promote the activation of CAFs,thus promoting cell proliferation and migration.The phenomenon may be regulated by the NF-?B pathway. |
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