Effect And Mechanism Of Ellagic Acid On The Proliferation And Apoptosis Of Human Pancreatic Carcinoma PANC-1 Cells

Objective To investigate the effect and mechanism of ellagic acid on the proliferation and apoptosis of human pancreatic carcinoma PANC-1 cells and provide the experimental basis for its application to the treatment of pancreatic carcinoma.Methods (1)The human pancreatic carcinoma PANC-1 cells were cultured in vitro for 24h,then intervened by different concentrations(0,2μg/ml,4μg/ml,6μg/ml,8μg/ml) of ellagic acid for 24h,48h and 72h.The morphological changes of cells were observed by inverted microscope and the inhibition rate of proliferation were assessed by MTT assay.(2)The human pancreatic carcinoma PANC-1 cells were cultured in vitro for 24h,then intervened by different concentrations(0,2μg/ml,4μg/ml,6ug/ml) of ellagic acid for 24h,48h and 72h:①the apoptotic morphological changes of cells were observed by Hoechst33258 staining.②the apoptosis rate of cells were detected through Annexin V-FITC/PI double staining.③the changes of cell cycle were detected by PI staining.④the expression of COX-2 and NF-KB(p65) were tested by Western blot.Results (1)There were more phenomena of pyknosis and karyorrhexis and more floating cells,but fewer adherent cells observed by inverted microscope when the PANC-1 cells were intervened by the higher concentration of ellagic acid for the longer time.Ellagic acid significantly inhibited the proliferation of the PANC-1 cells in a time- and dose-dependent manner compared with the negative groups(all P<0.05) revealed by MTT.(2)①after stained by Hoechst33258, the higher concentration of ellagic acid for the longer time,the more cells with pyknosis and karyorrhexis(apoptotic cells), but the fewer living cells compared with the negative groups(all P<0.05).②ellagic acid significantly induced the apoptosis in a time- and dose-dependent manner compared with the negative groups(all P<0.05) revealed by Annexin V-FITC/PI double staining.③ellagic acid could arrest the cell cycle in the Gl phase in a time- and dose-dependent manner compared with the negative groups(all P<0.05) revealed by PI staining.(5)Westem blot revealed that when the PANC-1 cells were intervened by the higher concentration of ellagic acid for the longer time,the expression of COX-2 and NF-κcB(p65) decreased more obviously.Conclusion The ellagic acid can significantly inhibit the proliferation and induce the apoptosis of the human pancreatic carcinoma PANC-1 cells in a time- and dose-dependent manner in vitro.The potential mechanism might to be related to arresting the cell cycle and decreasing the expression of COX-2 and NF-κB.