TPI1 Promotes Gastric Cancer Proliferation,Invasion And Cell Cycle Through Glycolysis And P53 Signaling Pathway

Objective: Triosephosphate Isomerase 1(TPI1)is a key enzyme in the process of cell energy metabolism.Abnormal changes of TPI1 expression level have been observed in a variety of malignant tumors and promote the malignant progression of tumors.However,research on TPI1 in gastric cancer(GC)is limited,and the mechanism of its action in GC is still unclear.This study intended to explore the biological role and potential molecular mechanism of TPI1 in GC.Methods:(1)The differentially expressed molecules in serum of GC patients were screened based on proteomics,and the expression level of TPI1 in cancer tissues and adjacent tissues was analyzed by bioinformatics.Furthermore,the possible mechanism was explored by GSEA enrichment analysis.We analyzed pathological data to determine the correlation between TPI1 expression and clinical characteristics using pathological data.(2)IHC with GC tissue microarray was used to detect TPI1 expression in the tissues of patients with GC,as well as the correlation between the expression level of TPI1 and clinical characteristics was further statistically analyzed in combination with patient data.(3)Western blot was used to detect the protein level of TPI1 in five GC cells,and determine the appropriate cell line for subsequent experimental research.The lentivirus was transfected to establish stable transgenic strains with high and low expression of TPI1.(4)Clonogenic assay,CCK-8 and Ed U were used to observe the effect of TPI1 on the proliferation of GC cells;the function of TPI1 on the invasion and metastasis of GC cells was tested using Transwell and scratch test.Flow cytometry can analyze the effect of TPI1 on cell cycle.CCK8 experimental detection of IC50 to investigate the effect of TPI1 on the sensitivity of oxaliplatin.(5)In nude mice,TPI1 was tested for its effect on subcutaneous tumorigenesis.(6)The effect of TPI1 on cell energy metabolism was detected by glycolysis pressure test.(7)The cell cycle related protein and p53 signaling pathway related protein were probed by Western blot to reveal the relevant mechanism of affecting the malignant progression of GC cells.Results:(1)The results of proteomics showed that seven genes including TPI1 were highly expressed in the serum of GC patients.The transcriptomics analysis showed that TPI1 was highly expressed in GC,and its high expression was significantly correlated with the age.(2)Immunohistochemical analysis showed that TPI1 was highly expressed in GC tissues,and was related to gender,depth of invasion and lymph node metastasis.(3)MKN45 and NCI-N87 were selected as the cell lines needed for the experiment.(4)The cell function test showed that after knockdown of TPI1,MKN45’s proliferation ability decreased,cell cycle was blocked.In addition,invasion and metastasis ability was inhibited,and sensitivity of oxaliplatin decreased.After overexpression of TPI1,the proliferation,invasion and metastasis of N87 cells were enhanced,the process of cell cycle was accelerated,and sensitivity was improved.(5)The subcutaneous tumorigenesis experiment in nude mice showed that the tumor mass and volume were significantly decreased after knockdown of TPI1,while the tumor mass and volume were significantly increased in the high expression of TPI1 group.(6)The energy metabolism experiment showed that the extracellular acidification rate of TPI1-knockdown cell was significantly decreased.(7)GSEA enrichment analysis showed that TPI1 was mainly enriched in p53 signaling pathway,cell cycle,glycolysis and gluconeogenesis,and Western blot showed that TPI1 was involved in cell cycle;TPI1 affects the malignant behavior of GC through p53 signaling pathway.Conclusion: There is an increase in TPI1 expression in GC,which is associated with the depth of invasion and lymph node metastasis.The high expression of TPI1 can promote the proliferation,invasion,and metastasis of GC,as well as accelerate its cell cycle.The high expression of TPI1 also enhanced cell glycolysis,making tumor cells have better ATP energy supply.In addition,we found that TPI1 is mainly involved in the malignant behavior of GC by inhibiting p53 signaling pathway.