| Objectives To investigate the effects of miR-192-5p on proliferation,migration,invasion and epithelial-mesenchymal transition(EMT)of pancreatic cancer by targeting ZEB2.The study will provide an experimental basis for specific targeted therapy of pancreatic cancer.Methods 1 The expression of miR-192-5p and ZEB2 were detected by qRT-PCR and Western blot in normal pancreatic ductal epithelial cells HPNE and pancreatic cancer cells PANC-1 and As PC-1.2 The effects of miR-192-5p mimics or inhibitor transfection on the proliferation,migration and invasion of PANC-1 and As PC-1 cells were analyzed by CCK-8,colony formation,wound healing and Transwell assays.3 The effects of miR-192-5p mimics or inhibitor transfection on the expression of EMT markers in PANC-1 and As PC-1 cells were determined by qRT-PCR and Western blot.4 Bioinformatic prediction website was used to predict the candidate target gene of miR-192-5p.Double luciferase reporter vector assay was employed to verify the targeting relationship between miR-192-5p and its target gene.5 The Starbase database was used to evaluate the correlation between the expression of miR-192-5p and ZEB2 in pancreatic cancer tissues.6 The effects of knockdown or overexpression of ZEB2 on the proliferation,migration,invasion and expression of EMT markers of PANC-1 and As PC-1 cells were detected by CCK-8,colony formation,wound healing,Transwell and immunofluorescence assay.7 CCK-8,Transwell and immunofluorescence assay were performed to detect whether ZEB2 could rescue the inhibitory effect of miR-192-5p on the proliferation,migration,invasion and EMT process in PANC-1 cells.Results 1 The results of qRT-PCR and Western blot assays showed that the expression of miR-192-5p was significantly decreased and the expression of m RNA and protein of ZEB2 was significantly increased in PANC-1 and As PC-1 cells,compared with that in HPNE cells.The differences were statistically significant(P<0.05).2 The results of CCK-8 showed that the proliferation of PANC-1 and As PC-1 cells transfected with miR-192-5p mimics was significantly decreased,compared with mimics NC group,while the proliferation of PANC-1 and As PC-1 cells transfected with miR-192-5p inhibitor was significantly enhanced,compared with inhibitor NC group.The differences were statistically significant(P<0.05).3 The results of colony formation assay showed that the colony formation ability of PANC-1 and As PC-1 cells transfected with miR-192-5p mimics was significantly lower than mimics NC group,while the colony formation ability of PANC-1 and As PC-1 cells transfected with miR-192-5p inhibitor was significantly higher than inhibitor NC group,and the differences were statistically significant(P<0.05).4 The results of wound healing and Transwell assay showed that the migration and invasion of PANC-1 and As PC-1 cells transfected with miR-192-5p mimics was significantly decreased,compared with mimics NC group.On the contrary,the migration and invasion ability of PANC-1 and As PC-1 cells transfected with miR-192-5p inhibitor was significantly enhanced,compared with inhibitor NC group.The differences were statistically significant(P<0.05).5 The results of bioinformatics prediction and double luciferase report experiment suggested that the luciferase activity in 293 T cells,PANC-1and As PC-1 cells significantly decreased after co-transfection of miR-192-5p mimics and ZEB2-3’UTR plasmids,compared with mimics NC and ZEB2-3’UTR plasmid cotransfection group,and the differences were statistically significant(P<0.05).However,there were no significant change in luciferase activity in 293 T cells,PANC-1 and As PC-1cells after co-transfection of miR-192-5p mimics and ZEB2-3’UTR-mut plasmids,compared with mimics NC and ZEB2-3’UTR-mut plasmid co-transfection group.6Starbase database analysis showed that there was a negative correlation between the expression of ZEB2 and miR-192-5p in pancreatic cancer tissues,and the difference was statistically significant(P<0.05).7 The results of CCK-8,colony formation,wound healing,Transwell,qRT-PCR,Western blot and immunofluorescence showed that knockdown ZEB2 inhibited the proliferation,migration,invasion and EMT process of PANC-1 and As PC-1 cells,compared with the si-NC group,while overexpression of ZEB2 promoted the proliferation,migration,invasion and EMT process of PANC-1 and As PC-1 cells,compared with the pc DNA3.1 control group.The differences were statistically significant(P<0.05).8 CCK-8,Transwell and immunofluorescence experiments revealed that overexpression of ZEB2 could restore the inhibitory effect of miR-192-5p on proliferation,migration,invasion and EMT process of PANC-1 cells,and the differences were statistically significant(P<0.05).Conclusions 1 Overexpression of miR-192-5p or knockdown of ZEB2 inhibited the proliferation,migration,invasion and EMT process of pancreatic cancer cells PANC-1 and As PC-1.2 Inhibition of miR-192-5p or overexpression of ZEB2 promoted the proliferation,migration,invasion and EMT process of pancreatic cancer cells PANC-1 and As PC-1.3miR-192-5p might inhibit the proliferation,migration,invasion and EMT process of pancreatic cancer cells by targeting ZEB2.Figure 25;Table 0;Reference 80... |
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