TWIST1 And ZIP4 Regulate The Role And Mechanism Of Emt In Invasion And Metastasis Of Pancreatic Cancer

Purposes:1.To explore the expression level,clinical significance of TWIST1 and ZIP4 in pancreatic cancer;2.To study the regulation of TWIST1 and ZIP4 on proliferation,invasion and migration of pancreatic cell lines PANC-1 and CFPAC-1 and the molecular mechanism of EMT-related proteins.Methods:1.A total of 81 cases of pancreatic cancer tissue paraffin specimens and 45 cases of normal pancreatic tissue paraffin specimens adjacent to cancer were collected from Pathology Department of The First Affiliated Hospital of Bengbu Medical College from July 2013 to May 2021.The expression levels of TWIST1,ZIP4 and EMT-related molecules（ZEB1）in pancreatic and normal pancreatic tissues were detected by Elivision ^TMimmunohistochemistry method.Chi-square test was used to analyze the relationship between the expression levels of relevant factors and clinicopathological parameters.Spearman correlation analysis was performed on the correlation between TWIST1,ZIP4 and ZEB1.Univariate Kaplan-Meier survival analysis and COX multivariate analysis were used to investigate the effect of expression and clinicopathological parameters on survival time of patients in TWIST1 and ZIP4;2.PANC-1 cell lines were used to transfect TWIST1 overexpressed plasmids and ZIP4overexpressed plasmids,CFPAC-1 cell lines were used to transfect si ZIP4.The transfection efficiency was observed by fluorescence microscopy.Western blot was used to verify the expression of TWIST1,ZIP4 and EMT-related proteins ZEB1 and E-cadherin.CCK-8 proliferation assay was used to detect the proliferation of tumor cells.The transwell chamber assay was used to verify the ability of tumor cells to invade and migrate.Results:1.The expression of TWIST1,ZIP4 and ZEB1 proteins in pancreatic cancer tissues was higher than that in normal pancreatic tissues（P<0.05）;TWIST1 and ZIP4expression levels were correlated with tumor metastasis and high TNM staging of pancreatic cancer（P<0.05）,ZEB1 expression level was correlated with tumor metastasis（P<0.05）;2.The expression levels of ZIP4,TWIST1 and ZEB1 proteins in pancreatic cancer tissues were pairwise correlated（P<0.05）.Survival analysis showed that overall survival was correlated with the degree of tumor differentiation,distant metastasis,TNM stage,ZIP4、TWIST1 and ZEB1 expression（P<0.05）.Moreover,the degree of tumor differentiation is an independent prognostic risk factor in patients with pancreatic cancer;3.After changing the expression levels of TWIST1 and ZIP4,the proliferation ability,invasion ability and migration ability of pancreatic cancer cells were changed,and the differences were statistically significant（P<0.01）;4.After the increase of TWIST1 protein expression,there was no significant change in ZIP4 protein expression level.In contrast,ZEB1 and TWIST1 protein levels were significantly increased after high expression of ZIP4 protein,while E-cadherin protein levels were decreased（P<0.01）.After ZIP4 expression was knockdown,ZEB1 and TWIST1 protein expression levels were significantly decreased,while E-cadherin protein level was increased（P<0.01）.Conclusions:1.TWIST1,ZIP4 and ZEB1 are highly expressed in pancreatic cancer tissues and are associated with survival and prognosis of patients,which has certain clinical significance for prognosis assessment and targeted therapy of patients with pancreatic cancer;2.TWIST1 and ZIP4 play a role in the proliferation,invasion and migration of pancreatic cancer cells by regulating EMT-related proteins ZEB1;3.Changes in ZIP4 expression level could affect the expression of ZEB1 and TWIST1,and there may be some regulatory mechanism between them.