Clinical Significance Of Pan-cancer Biomarker Thymosin β10 In Prostate Cancer And Its Correlation With Hypoxia And Immune Microenvironment Regulation

Background:Thymosin β10(TMSB10)is believed to be upregulated in multiple cancers and can promote the development of various cancers,but the conclusions are still contradictory or controversial.The role of TMSB10 in cancer as a promoter or inhibitor remains unclear.Methods:In this study,transcriptional data,mutation information,tumor grading,and prognosis information of cancer tissues/paracancer tissues from 33 cancer types were downloaded from The Cancer Genome Atlas and GEPIA database.Non-paired/paired t-tests and analysis of Variance were used to analyze the differential expression of TMSB10 in cancer tissues and paracancer tissues in pan-cancers and in different tumor stages.Kaplan-Meier curves and univariate Cox regression analysis were used to evaluate the prognostic value of high-and low-TMSB10 expression in terms of overall survival(OS),disease-specific survival(DSS),disease-free interval(DFI),and progression-free interval(PFI)as tumor follow-up endpoints.Finally,Spearman correlation analysis was used to explore the correlation between TMSB10 expression and tumor mutational burden(TMB)and microsatellite instability(MSI).Results:TMSB10 was found to be upregulated in most cancer tissues,especially adenocarcinoma and squamous cell carcinoma,while it was downregulated in renal clear cell carcinoma,acute myeloid leukemia,and low-grade glioma.Only in adrenocortical carcinoma,renal clear cell carcinoma,lung adenocarcinoma,prostate cancer,and thyroid cancer,TMSB10 expression was significantly increased with tumor staging.TMSB10 expression was positively correlated with TMB and MSI in breast cancer,colon cancer,lung adenocarcinoma,and prostate cancer.Survival analysis showed that TMSB10 overexpression was a poor prognostic factor for OS and DSS in adrenocortical carcinoma,renal clear cell carcinoma,low-grade glioma,hepatocellular carcinoma,and lung adenocarcinoma,but a protective factor in bladder cancer,ovarian cancer,and melanoma.From the perspective of DFI and PFI,high TMSB10 expression showed excellent predictive value for the progression of prostate adenocarcinoma.Conclusions:TMSB10 is upregulated in most cancers,especially adenocarcinoma and squamous cell carcinoma,but its overexpression does not necessarily indicate higher tumor staging or worse prognosis.Based on all the analyzes,more evidence showed that TMSB10 is a potential prognostic biomarker with great research value for predicting poor outcomes in adrenocortical carcinoma,lung adenocarcinoma,and prostate adenocarcinoma.Background:Due to the heterogeneity of prostate and the complexity of treatment,prostate cancer inevitably develops resistance to androgen deprivation therapy and progresses to metastasis,necessitating the exploration of new therapeutic targets and prognostic markers.The previous study suggested that Thymosin β10(TMSB10)is a potential prognostic marker for prostate cancer,but its expression and pathogenic mechanism in prostate cancer have not been reported.Methods:This study utilized multiple prostate cancer cohorts,including The Cancer Genome Atlas(TCGA),the International Cancer Genome Consortium,and the cohorts of Gene Expression Omnibus,to investigate the prognostic value of high-and low-TMSB10 expression in terms of overall survival(OS),siochemical recurrence(BCR),and progression-free interval(PFI)using survival analyses such as KaplanMeier curves and univariate/multivariate Cox regression.Additionally,the clinical characteristics differences between high-and low-TMSB10 expression groups was analyzed.Authors further explored TMSB10 expression in different prostate cancer risk groups,treatment status,and disease stages.Gene Set Enrichment Analysis(GSEA)was performed on eight prostate cancer cell lines in the Cancer Cell Line Encyclopedia database and TCGA prostate cancer cohort data.Immunohistochemistry,CCK-8,and Transwell experiments were conducted to investigate the correlation between TMSB10 expression and prostate cancer staging and its effect on the proliferation,migration,and invasion of prostate cancer LNCaP and DU 145 cells after interfering with TMSB10 expression in vitro.Results:Based on survival analysis and meta-analysis of multiple cohorts and pooled cohorts,high TMSB10 expression was found to be associated with poor OS,BCR,and PFI,and it was an independent predictor for BCR and PFI.In the high TMSB10 group,clinical features such as PSA,Gleason score,T stage,tumor metastasis,and positive surgical margins were significantly increased in prostate cancer patients.Functional analysis showed that TMSB10 expression was closely related to the epithelial-mesenchymal transition(EMT)pathway and the conversion of androgenindependent phenotypes of prostate cancer cells,and that hypoxia-related genes,immune-related components,and regulatory pathways were activated in the high TMSB10 group.Immunohistochemistry experiments showed that TMSB10 staining increased with tumor stage and disease progression.In vitro experiments showed that interference with TMSB10 expression led to decreased proliferation and significantly reduced migration and invasion of LNCaP and DU 145 cells in continuous culture using CCK-8 assays and Transwell assays.Conclusions:TMSB10 is a potential prognostic factor for prostate cancer,and its expression increases with tumor staging and disease progression.Its oncogenic mechanism may be related to the EMT pathway and androgen-independent phenotype transformation,and high TMSB10 expression is closely related to hypoxia and the immune microenvironment.Interfering with TMSB10 expression can significantly inhibit the proliferation,migration and invasion ability of LNCaP and DU 145 cells.Background:Hypoxia is a characteristic of tumor development,and researchs has demonstrated varying degrees of hypoxic microenvironments in prostate cancer.The previous study have shown that thymosin β10(TMSB10)overexpression was closely related to hypoxia microenvironment in prostate cancer,but it is unclear how they regulate each other and whether they affect the prognosis of prostate cancer.Methods:This research first used Spearman correlation analysis to analyze the correlation between TMSB10 and hypoxia-inducible factor-1α(HIF-1α)in two Gene Expression Omnibus(GEO)datasets.Then,authors used single sample Gene Set Enrichment Analysis(ssGSEA)and univariate Cox regression to calculate the hypoxia pathway scores of patients in The Cancer Genome Atlas(TCGA)prostate cancer cohort and their relationship with prognosis.Next,study established a hypoxia risk score model using algorithms such as protein interaction networks,univariate Cox regression,LASSO regression,and multivariate Cox regression,with the TCGA prostate cancer cohort as the training set and the GEO GSE116918 cohort as the test set.After establishing the model,study validated its applicability and ability to distinguish hypoxic microenvironments in both the training and test sets.Then,the expression of TMSB10 in the high hypoxia score group was analyzed.Finally,authors cultured LNCaP and DU 145 cell lines in hypoxic and normoxic environments and tested the changes in TMSB10 expression in both groups.Results:Spearman correlation analysis showed that TMSB10 and HIF-1α were coexpressed in prostate cancer.ssGSEA and univariate Cox regression analysis confirmed the presence of a hypoxic microenvironment in prostate cancer,which was a risk factor for poor prognosis.Through a series of analyses and calculations,a prostate cancer specific hypoxia risk score model consisting of seven genes prostate was established.The Kaplan-Meier curves and receiver operating characteristic curves of the training and test sets showed that the model had good applicability.Principal component analysis and t-distributed stochastic neighbor embedding analysis showed that the gene expression patterns of the high and low hypoxia risk score groups were different,and GSEA confirmed significant enrichment of hypoxia-related genes in the high hypoxia risk score group.In the high hypoxia risk score group,and A549,PC3 cells cultured under hypoxia in public datasets,TMSB10 expression was significantly upregulated.Further cell experiments showed that TMSB10 expression in LNCaP and DU145 cells initially decreased within the first 24 hours of hypoxic culture,and then gradually increased and surpassed the normoxic group.Conclusions:Different degrees of hypoxic microenvironment exist in prostate cancer.TMSB10 is co-expressed with HIF-1α.The established prostate cancer specific hypoxia risk score model has good discriminative ability for poor prognosis and tissue hypoxia status.The upregulation of TMSB 10 expression in the high hypoxia risk score group and cell experiments under hypoxic culture conditions suggest that hypoxia has a regulatory effect on prostate cancer cells and that TMSB 10 is upregulated in a tissue hypoxic environment.Background:Immunotherapy for prostate cancer is being considered as a treatment option for patients who have developed resistance to existing therapies or have castration-resistant prostate cancer.However,due to its patients selectivity,new biomarkers are needed to be searched for screening patients who are sensitive to immunotherapy.The second part of the study found that high expression of Thymosinβ10(TMSB10)is closely related to immune regulation,but its specific role in the immune microenvironment remains unknown.Methods:Firstly,authors analyzed the immune infiltration of high and low TMSB10 expression groups based on transcriptome matrix of The Cancer Genome Atlas(TCGA)prostate cancer cohort using four algorithms including CIBERSORT,single sample Gene Sets Enrichment Analysis(ssGSEA),Immune Cell Abundance Identifier(ImmuCellAI),and Immunophenoscore(IPS).Next,authors tested the expression of immune checkpoint after TMSB10 interference in vitro and verified it using immune fluorescence in prostate cancer biopsy tissues.Finally,authors analyzed the response to immunotherapy in different TMSB10 expression groups based on the IMvigor210 dataset and IPS scores for different immune phenotypes of prostate cancer patients in The Cancer Immunome Atlas.Results:Overall immune activation,including immune suppression,was found in the high TMSB10 expression group.CD8+T cells,dendritic cells,regulatory T cells,and immune checkpoints were activated in the high TMSB10 expression group.ESTIMATE analysis showed a positive correlation between TMSB10 expression and immune and stromal scores.Moreover,genes related to myeloid-derived suppressor cells and Treg cells were positively correlated with TMSB10 expression.In addition,high expression of immune checkpoints,such as programmed cell death ligand(PD-L)1,PD-L2,CD276,and indoleamine 2,3-dioxygenase(IDO1),was detected in the high TMSB 10 expression group,which decreased after TMSB 10 interference in vitro.The fluorescence intensity of these check points was significantly enhanced in the high TMSB 10 expression group in tissue immunofluorescence.With regard to the immune response analysis,TMSB 10 expression was found to predict the response to PD-L 1/PD1/CTLA4 immunotherapy,and high TMSB 10 expression was associated with increased sensitivity to these immunotherapies.Conclusions:High TMSB10 expression was associated with both immune activation and immune suppression,possibly due to immune exhaustion primarily involving CD8+T cells.High TMSB10 expression was positively correlated with the enrichment of immune-suppressive cells and the co-expression of immune checkpoints.Additionally,high TMSB 10 expression was found to be a potential biomarker for predicting the response to immunotherapy,as it was associated with increased sensitivity to these treatments.