FTO Promotes The Proliferation,migration And Invasion Of Pancreatic Cancer Cells By Regulating The Glycolysis Related Gene PFKM

Objective: The main purpose of this paper is to explore whether FTO could affect the glycolysis process of pancreatic cancer cells by regulating the glycolysis related gene,such as phosphofructokinase(PFK),hexokinase(HK),pyruvate kinase(PK),phosphoglycerate mutase(PGAM),lactate dehydrogenase(LDH)to promote the proliferation,migration and invasion of pancreatic cancer.Methods: TCGA database and GTEx database were used for gene expression and correlation analysis.The changes of gene m RNA and protein levels were verified by RTqPCR and Western blot.Transfect cells with siRNA to obtain cells that knock down the target gene instantaneously.Using lentivirus vector to construct stable knock down cell line.Overexpression plasmid was used to increase the target gene PFKM.CCK8 assay and colony formation were used to determine the level of cell proliferation.Wound healing assay and Transwell cell assay were used to detected the change of cell migration and invasion.Glucose consumption and lactate production were used to detect the changes of cell glycolysis level.Results: In this paper,we verified the expression of FTO in four pancreatic cancer cell lines.Compared with normal pancreatic duct epithelial cells,FTO is highly expressed.Successfully constructed a stable FTO knock down cell line.Four functional experiments confirmed that knockdown of FTO could inhibit the function of pancreatic cancer cells,such as inhibiting the ability of cell proliferation,migration and invasion.By analyzing the correlation between FTO and glycolysis related genes used GEPIA2 website,the results showed that FTO was strongly correlated with PFKM(R=0.9).Further research found that the m RNA and protein levels of PFKM decreased significantly after knocking down FTO.After knocking down PFKM with siRNA,it was observed that the proliferation,migration and invasion of pancreatic cancer cells decreased,and glucose consumption and lactate production decreased.It was found that the glucose consumption and lactate production of pancreatic cancer cells were also reduced after FTO knock down.In rescue experiment,overexpression of PFKM could restore the decline of pancreatic cancer cell function caused by down-regulation of FTO partially.Conclusion: In pancreatic cancer,FTO promotes the glycolysis of pancreatic cancer cells by regulating PFKM,and promotes the proliferation,migration and invasion of pancreatic cancer cells.