Research Of The Role Of FTO In Thyroid Papillary Carcinoma

Objective:The fat mass-and obesity-associated enzyme(FTO)was M6 A demethylase which was first discovered.This study aims to clarify the expression and function of FTO in thyroid papillary carcinoma(PTC),and analyze the mechanism of progression of thyroid papillary carcinoma with FTO at the metabolic level.Methods:At first,we explored the expression level of FTO in PTC cancer tissues and paracancerous tissues by immunohistochemistry and Western blotting.Thyroid papillary cancer cell strain(TPC-1,BCPAP)with FTOi Gene stable silencing was established by infection with lentivirus.The proliferation ability of TPC-1 and BCPAP with FTO knockdown was detected by CCK8 and colony-forming unit assays.Apoptosis was analyzed by flow cytometry.Migrated ability and invasive ability of TPC-1 and BCPAP cells with FTO knockdown were determined with Wound-Healing Assay、Transwell and EMT-related proteins assay.TPC-1 in the control group(CON77)and knockdown group(FTOi)was tested by Metabolomics with High Performance Liquid Chromatography-Quadrupole Time-of-Flight Tandem Mass Spectrometry(HPLC-Q-TOF-MS-MS),to screen differential metabolite and pathways.Results:1.FTO was highly expressed in PTC by Immunohistochemistry and Western blotting(p < 0.05).2.The proliferation ability of TPC-1 and BCPAP knocked down by FTO gene was significantly reduced(p < 0.05).3.The level of apoptosis of TPC-1 and BCPAP with FTO gene knockdown increased by flow cytometry(p < 0.05).4.Invasion and migration of TPC-1 and BCPAP with FTO knockdown Impaired significantly(p<0.05),while N-cadherin and Vimentin increased,whose gray values is statistically significant,p<0.05.5.By processing original data of metabolomic,we found significant variation in metabolic profiling between group CON77 and FTOi in TPC-1.By combining VIP >1.0 and FC >1.5 with p<0.05,58 potential differential metabolites were screened out,including taurodeoxycholic acid,levetiracetam,arginine,isoamylcarnitine,oxalic acid,glycerophosphate choline,Tika carnitine,sterol,estradiol and glibenclamide.Among them,40 metabolites,such as taurodeoxycholic acid,levetiracetam,arginine,isoamylcarnitine,oxalic acid,tika carnitine,sterol,estradiol,glibenclamide and glycerophosphate choline,were up-regulated,and 18 metabolites,such as etoposterone,2-dimethylfuran,isovalerylglycine,pentoxifylline,benzylamine,fluorobenzoylpropionic acid,imiquimod,toluenesulfonamide,oleoacylcarnitine,gentilic acid and D-ribose5-phosphate,were down-regulated in TPC-1 with FTO gene silencing.At the same time,6 different metabolic pathways related to FTO gene silencing were screened out in TPC-1 cells,respectively: glycerol phospholipid metabolic pathway,cysteine and methionine metabolic pathway,pyrimidine metabolic pathway,pyruvate metabolic pathway(glycolysis/gluconeogenesis),sulfur metabolism and steroid biosynthesis.Conclusion:FTO,which is highly expressed in PTC,mediates the proliferation,invasion,and metastasis,and inhibits the apoptosis of PTC cell.Which may be related to lipid metabolism,amino acid metabolism,glycolysis and pyrimidine metabolism.