Study On The Role Of Human Peptide Deformylase In Colorectal Cancer

Background: Colorectal cancer seriously endangers human life and health,and lacks effective treatment drugs.Therefore,it is urgent to clarify the pathogenesis of colorectal cancer and develop effective treatment strategies.Finding new targets for colorectal cancer will provide a theoretical basis for the development of new targeted drugs for colorectal cancer.Human peptide deformylase(hsPDF)mainly exists in mitochondria and plays an important role in mitochondrial protein synthesis by catalyzing deformylation.It has been found that hsPDF is highly expressed in a variety of tumors,especially in colorectal cancer.Our previous studies also showed that hsPDF inhibitors have good in vivo and in vitro anti-proliferative activity against colorectal cancer,suggesting that hsPDF may become a new target for anti-colorectal cancer,but its role in the development of colorectal cancer remains unclear.Objective: To study the relationship between hsPDF expression and proliferation,migration and cell cycle of colorectal cancer cells,exploring the possible mechanism of hsPDF action,to provide evidence for hsPDF as a new target of anti-colorectal cancer drugs.Methods:(1)Retrieving the database of gene expression of tumors and searching the expression of hsPDF gene in various tumors and adjacent tissues.(2)Real time PCR and Western blot were used to detect the difference of hsPDF gene and protein expression between normal cells and colorectal cancer cells,colorectal cancer tissues and adjacent tissues.(3)Human colorectal cancer HCT116 cell line and control cell line with stable overexpression or inhibited expression were constructed by using lentivirus vector system.(4)CCK-8 and colony formation assay were used to detect the effect of hsPDF overexpression on the proliferation of colorectal cancer HCT116 cells.(5)Establishment of xenograft model in nude mice to detect the effect of hsPDF overexpression on the proliferation of HCT116 cells and the anti-tumor activity of hsPDF inhibitor PDF64 in vivo.(6)The relationship between hsPDF expression and HCT116 cell migration was detected by cell scratch assay.(7)Western blot and ATP detection kit were used to detect the effect of hsPDF overexpression on mitochondrial protein expression and ATP production.(8)Flow cytometry and Western blot were used to determine the effect of hsPDF overexpression on the cell cycle of HCT116.(9)The effect of hsPDF on the activation of AKT/m-TOR signaling pathway and the expression of cyclin in colorectal cancer cells and animal tumors was determined by Western blot.Results:(1)By searching the Gepia(http://gepia.cancer-pku.cn/)tumor gene expression database,it was found that hsPDF gene was highly expressed in many types of tumors,especially in colorectal and lymphoid cancers.(2)In colorectal cancer cells and tissues,the levels of hsPDF m RNA and protein were significantly increased compared with normal cells and adjacent tissues,which was consistent with the results of database query.(3)In stable cell lines,overexpression of hsPDF promotes the proliferation of HCT116 cells.On the contrary,downregulation of hsPDF expression inhibits the proliferation of HCT116 cells and HT-29 cells.(4)Similar results were obtained in animal experiments: hsPDF overexpression significantly promoted the proliferation of HCT116 tumor cells in vivo,while hsPDF inhibitor PDF64 significantly inhibited the growth of HL60 transplanted tumors in vivo.(5)Overexpression of hsPDF induced a significant increase in mitochondrial protein expression and ATP production in HCT116 cells.(6)Overexpression of hsPDF promotes the production of cyclin in HCT116 cells,accelerates cell cycle progression and accelerates cell migration.(7)The phosphorylation level of AKT/m-TOR signaling pathway in hsPDF overexpressed HCT116 cells and tumor tissues was significantly increased,which was contrary to the effect of PDF64 on this pathway.Conclusion: The expression of hsPDF was positively correlated with the proliferation of HCT116 colorectal cancer cells.Upregulation of hsPDF expression can promote the proliferation and migration of HCT116 tumor cells,accelerate cell cycle,affect mitochondrial function,and activate AKT/m-TOR pathway,which may be the mechanism of regulating the occurrence and development of colorectal cancer.This study provides a theoretical basis for revealing the biological function of hsPDF and confirming its status as a new target of anti-colorectal cancer.