MIF Promotes The Migration,Invasion And Proliferation Of Pancreatic Cancer Cells By Regulating ILF3 Through HNRNPA2B1

Objective:Pancreatic cancer is a great threat to life due to its relatively late detection and high degree of malignancy.It is very important to explore the mechanism of its occurrence and development for the early detection and early treatment of pancreatic cancer.Macrophage migration inhibitory factor(MIF)is a cytokine highly expressed in tumor microenvironment and closely related to tumor function.MIF is highly expressed in pancreatic cancer and is closely related to the occurrence and development of pancreatic cancer.m~6A methylation is one of the main forms of post-transcriptional regulation and affects the occurrence and development of tumors.This process involves the participation of methyltransferase,demethylase,reader and other related proteins.Heterogeneous nuclear ribonucleoprotein A2B1(HNRNPA2B1)is a reading protein.Interleukin enhancer binding factor 3(ILF3),also known as NF90/NF110,exerts different functions such as stabilizing m RNA and inhibiting translation by binding to m RNA.Recently,the role of ILF3 in cancers such as digestive tract cancer and breast cancer has been gradually revealed.At present,it is not completely clear how MIF affects the function of cancer cells in pancreatic cancer.In this paper,the correlation between the expression of MIF,HNRNPA2B1 and ILF3 in pancreatic cancer and the function of pancreatic cancer cells were investigated by conventional experimental methods.Methods:Construct a lentivirus knockdown MIF stable cell line to verify how it affects the function of pancreatic cancer cells.The expression differences of MIF and m~6A methylation-related proteins in pancreatic cancer and adjacent tissues(normal tissues)were analyzed by bioinformatics,and the correlation between them was analyzed to determine the MIF-related m~6A methylation-related proteins that need to be studied.String database was used to find proteins related to m~6A methylation-related protein expression.Lentiviral transfection to construct stable cell lines;the transfection effect was verified by q RT-PCR and Western blotting(WB).The effects of MIF knockdown,HNRNPA2B1knockdown and overexpression,ILF3 knockdown and overexpression on the function of pancreatic cancer cells were verified by CCK8 assay,cell scratch assay,Transwell chamber migration and invasion assay.\Results:GEPIA2 analysis showed that MIF was differentially expressed and correlated with multiple m~6A methylation-related genes(METTLE16,ZC3H13,HNRNPA2B1,etc.)between pancreatic cancer and adjacent tissues.PCR screening results after lentivirus knockdown of MIF suggested that HNRNPA2B1 decreased most significantly.WB results verified that HNRNPA2B1 protein expression decreased after MIF knockdown.Knockdown of MIF could significantly inhibit the proliferation,invasion and migration of pancreatic cancer cells,but overexpression of HNRNPA2B1 on the basis of MIF knockdown could restore the function of pancreatic cancer cells.The String database and GEPIA2 database showed that there was a correlation between HNRNPA2B1 and ILF3 expression.The expression of ILF3 decreased significantly after knockdown of HNRNPA2B1 by sh RNA,and the function of pancreatic cancer cells was inhibited after knockdown of HNRNPA2B1 and ILF3,while the inhibition was restored after overexpression of ILF3 in the HNRNPA2B1 knockdown cell line.Conclusion:MIF regulates the expression of ILF3 through HNRNPA2B1 in pancreatic cancer,thereby affecting the proliferation,invasion and migration of pancreatic cancer cells.