The Mechanism Of SATB1 Regulating The Expression Of CXCL1 And CXCL8 Involved In The Growth And Progression Of Pancreatic Cancer

Objective:Pancreatic cancer has become the poorest prognostic digestive system tumor due to its rapid progress and metastasis.The overexpression of SATB1 has been found in many solid tumors,and to be associated with the progression of cancers.Studies have shown that it induces cancer cell invasion and metastasis by regulating downstream gene expressions.Members of the CXC chemokine family,chemokine-1(CXCL1)and chemokine-8(CXCL8),are known to be involved in tumorigenesis and angiogenesis in various tumors through promoting cell proliferation,invasion and metastasis.This article will explore the role of CXCL1 and CXCL8 in growth and progression of pancreatic cancer,which provides a theoretical basis for the application of chemokines in the diagnosis and treatment of pancreatic cancer.Methods:1.Microarray technology and Venn analysis were applied to find out the target gene regulated by SATB1,and the effect of SATB1 on tartget gene was determined by Real-time PCR.GEPIA online database was applied to analyse the expression of CXCL1?8 in patients with pancreatic cancer and adjacent tissue,the relationship between the expression level of CXCL1.8 with the pathological stage and the prognosis of patients in pancreatic cancer.2.Short hair RNA(shRNA)was used to knock down CXCL1 and CXCL8.And the pancreatic cancer cell lines with high interference efficiency at sites 3 and 4 were chosen for the experiments.The proliferation,migration,and adhesion ability of pancreatic cancer cells were detected by in vitro functional tests such as MTT,clone assay,wound healing assay,and adhesion assay.3.Nude mice tumor xenograft model was made by subcutaneous injection of pancreatic cancer cell lines.Ki-67 and CD34 expression in pancreatic cancer was tested by immunohistochemistry.4.The tumor conditioned medium(TCM)was prepared by the supernatant of pancreatic cancer cells of site 3 and 4.HUVECs were treated with the TCM.The proliferation and migration ability of HUVECs were tested by MTT and wound healing assay.5.Microarray technology and bioinformatics analysis were used to find out the DEGs and detect the potential mechanism.Results:1.SATB1 up-regualtes the expression of CXCL1 and CXCL8.The expression of CXCL1?8 in pancreatic cancer patients is significantly higher compared with the normal tissue and related with the pathological stage.The disease-free survival time of pancreatic cancer patients with high expression of CXCL8 was significantly decreased,while the disease-free survival time had no significant change in pancreatic cancer patients with high expression of CXCL1.2.In vivo and in vitro experiments showed that the interference of the expression of CXCL1 and CXCL8 in pancreatic cancer cells reduces the proliferation and migration ability of pancreatic cancer cells.3.The expression of Ki67 and CD34 was decreased through Immunohistochemical,suggesting that interference with CXCL1 and CXCL8 affected the tumorigenesis and angiogenesis of pancreatic cancer.4.MTT and wound healing assay showed that the proliferation and migration ability of HUVECs were inhibited by treated with TCM.5.The Microarray technology found out the DEGs between that interfered with CXCL1 expression group and control group,interfered with CXCL8 expression group and overexpressed CXCL8 group in HUVECs.GO analysis showed that the DEGs were mainly concentrated in signal transduction,cell communication,cell growth and apoptosis,and metabolism.KEGG pathway enrichment analysis showed that the DEGs are enriched in signal pathways such as PI3k-Akt,cGMP-PKG,and MAPK.PPI network indentified many role genes,up-regulated gene RBBP7 and down-regulated gene YBX1 were related with CXCL1.The up-regulated genes CD44,FOXM1,GATA3,the down-regulated genes CXCL10 were related with CXCL8.Conclusion:SATB1 promotes the proliferation and migration ability of pancreatic cancer cells by up-regulating the expression of downstream gene CXCL1 and CXCL8,and regulates the angiogenesis of pancreatic cancer,thereby promoting the growth and progression of pancreatic canecer.This result provides a certain reference for the chemokines CXCL1 and CXCL8 in the clinical application of pancreatic cancer.