| Objective: The research was to research the subcellular localization of denticleless protein homolog(DTL)and explore the possible biological role of DTL.Besides,based on bioinformatics analysis and combined with tissue and cytology experiments,the potential value of DTL expression in cancer diagnosis,prognosis and treatment prediction was studied and verified,and explored its mechanism through experiments.Methods: In our study,we detected the subcellular localization and expression of DTL by immunofluorescence(IF)and immunohistochemistry(IHC).Cell fraction extraction and western blot(WB)in a variety of cells and tissues were used to verify.Combined with GTEx,TCGA,TIMER and other open databases to obtain clinical follow-up documents and genome expression data,KEGG,R version 4.1.2,Kaplan Meier Plotter,Cytoscape and other tools were used to analyze expression,mutation,prognosis analysis,immune infiltration analysis,immunotherapy analysis,drug treatment target analysis and clinical pathological characteristics analysis of DTL.Gene set enrichment analysis(GSEA)and Gene Ontology(GO)enrichment analysis between groups with high and low DTL expression levels was performed.In addition,through the analysis of IF and IHC experimental results and clinical data,the prognosis,correlation of clinicopathologic characteristics and immune infiltration of hepatocellular carcinoma(LIHC)、 bladder urothelial carcinoma(BLCA)and stomach adenocarcinoma(STAD)were verified.Finally,by constructing DTL overexpression cell lines,flow cytometric(FC),live cell imaging,IF,WB and chromosome preparation were used to detect the changes in cell biological behavior caused by DTL overexpression.Results: It was confirmed that DTL had obvious subcellular translocation in the cell cycle,and different subcellular localization in different cells and different tissues.In addition,we have analyzed the bioinformatics and experimental results,and discovered that the expression of DTL in different cells and tissues was also significantly different.DTL was widely expressed in various cells and tissues,while it was overexpressed in tumor tissues except acute myeloid leukemia(LAML).Gene mutation analysis revealed that DTL mutation may not be the main cause of tumorigenesis due to its low mutation frequency in most tumors.Pan-cancer bioinformatics analysis showed that the expression of DTL was correlated with the prognosis,immune infiltration,therapeutic prediction,and clinical indexes.In addition,GSEA analysis showed that DTL was enriched in oocyte meiosis,pyrimidine metabolism,the cell cycle,the G2 M checkpoint,m TORC1 signaling and E2 F targets.It was verified that the overexpression of DTL was associated with immune cell infiltration and clinical indexes in LIHC,BLCA and STAD in our study.Furthermore,it was confirmed that the overexpression of DTL could regulate the cell cycle,promote cell proliferation and cause genomic instability in cultured cells,which may be the reason why DTL plays a role in the occurrence and progression of cancer.Conclusions:1.DTL had obvious subcellular translocation in the cell cycle,and different subcellular localization in different cells and different tissues.2.The expression level of DTL in cancer cells and tissues was increased,and was negatively related to the prognosis of most cancers,which may become a potential biomarker for predicting the prognosis of cancer.3.The expression level of DTL was positively correlated with markers of benefit from immune checkpoint blockade(ICB)in most cancers,which may become a potential biomarker for predicting the response of immunotherapy in cancer.4.The proliferation of DTL overexpression cell lines is accelerated,and the S phase is increased.DTL may lead to poor prognosis by promoting tumor growth and invasion through promoting cancer cell proliferation.5.With overexpression of DTL,DNA damage and apoptosis increased in cell lines and chromosome breaks became more frequent,which may cause genomic instability and result in poor prognosis and influencing immunotherapy. |
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