| Background:Pancreatic adenocarcinoma is a clinically common malignant tumor of the digestive system with a very poor prognosis with a five-year survival rate of only 8%in most patients.ANTRX1 is a highly conserved type I transmembrane glycoprotein found on the surface of cell membranes that is specifically expressed in tumor vessels and in some tumor cells themselves.Several studies have shown that ANTXR1 is a potentially excellent anti-tumor target,but there is no report on the expression and effects of ANTXR1 in pancreatic adenocarcinoma and its related mechanisms.Objective:(1)To clarify the expression of ANTXR1 in Pancreatic adenocarcinoma cell lines,normal pancreatic duct endothelial cells,pancreatic adenocarcinoma tumor tissue and adjacent tissues,and its relationship with clinicopathological parameters and prognosis.(2)To clarify the effect of ANTXR1 on the proliferation,invasion and metastasis of pancreatic adenocarcinoma cells.(3)To identify the possible mechanisms by which ANTXR1 affects the ability of pancreatic adenocarcinoma cells to proliferate,invade and metastasize.Methods:The expression of ANTXR1 in pancreatic adenocarcinoma cell lines and normal pancreatic ductal epithelial cells was detected by western-blot and qRT-PCR,and the expression of ANTXR1 in tumor tissues and adjacent tissues of pancreatic adenocarcinoma was detected by immunohistochemical staining.The relationship between ANTXR1 and clinicopathological parameters and prognosis was analyzed with clinicopathological data.Lentiviral infection constructs stable transgenic ANTXR1 knockdown cell lines,CCK-8 assay detects cell proliferation,flow cytometry detects apoptosis,Transwell assay detects invasion,scratch assay detects migration,Western-blot detects EMT-associated protein expression,nude mice subcutaneous tumor and in situ tumor assay validates the effect of ANTXR1 knockdown on pancreatic adenocarcinoma growth in vivo.Immunohistochemical staining was used to analyze the tumor and its adjacent tissues in nude mice.Transcription sequencing and bioinformatics were used to analyze the ANTXR1-related signaling pathways,qRT-PCR and Western-blot to verify the related pathway proteins.Results:The expression of ANTXR1 in pancreatic adenocarcinoma cells was significantly higher than that of normal pancreatic ductal epithelial cells by western-blot and qRT-PCR,and the high expression of ANTXRl in pancreatic adenocarcinoma tumor tissue was significantly higher than that of adjacent tissue by immunohistochemical staining(P<0.01).High expression of ANTXR1 was associated with higher TNM staging,median overall survival(OS)was 10.5 months(P<0.01)in patients with high expression of ANTXR1 and 13.0 months(P<0.01)in patients with low expression.Univariate and multivariate analyses suggested that ANTXR1 was an independent risk factor for prognosis in patients with pancreatic adenocarcinoma.Kaplan-Meier analysis revealed that the median overall survival of ANTXRl-high and preoperative serum CA19-9>270 U/mL group was 6.7 months;the median overall survival of ANTXRl-high or CA19-9>270 U/mL group was 10.7 months;the median overall survival of ANTXR1-low and CA19-9<270 U/mL group was 17.3 months(P<0.01).After ANTXR1 knockdown,pancreatic cancer cell proliferation decreased,apoptosis increased,invasion decreased,migration decreased,and EMT related phenotype was reversed.Transplant tumor growth slowed down in both subcutaneous and in situ tumor models of nude mice.Immunohistochemical staining showed that the expression of related markers was consistent with in vitro experiments.The molecular mechanism may be related to the ability of ANTXR1 to regulate the proliferation,invasion and metastasis of pancreatic adenocarcinoma through Wnt/P-catenin signaling pathway.Conclusion:The high expression of ANTXR1 in pancreatic adenocarcinoma is an independent risk factor for the prognosis of pancreatic adenocarcinoma patients.The combination of preoperative CA19-9 level and ANTXR1 indicate poor prognosis of pancreatic adenocarcinoma patients.ANTXR1 can promote the proliferation,invasion and metastasis of pancreatic adenocarcinoma by regulating the Wnt/?-catenin signaling pathway. |
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