Objective To identify pyroptosis,apoptosis,and necroptosis(PANoptosis)-related genes(PRGs)in clear cell renal cell carcinoma(ccRCC)for patient stratification and prognosis prediction.Methods We used differential expre...Objective To identify pyroptosis,apoptosis,and necroptosis(PANoptosis)-related genes(PRGs)in clear cell renal cell carcinoma(ccRCC)for patient stratification and prognosis prediction.Methods We used differential expression analysis and weighted gene co-expression network analysis(WGCNA)to identify ccRCC-specific PRGs.A prognostic model,the PANoptosis-index(PANI),was constructed using least absolute shrinkage and selection operator(LASSO)and Cox regression.The PANI model,comprising PRGs,was validated through single-cell RNA-sequencing(scRNA-seq),immunohistochemistry,and reverse transcription-quantitative polymerase chain reaction(RT-qPCR).Patient cohorts were categorized into high-and low-PANI groups,and the model’s performance was appraised using various metrics.External validation was performed with the E-MTAB-1980 dataset.Functional and gene set enrichment analyses distinguished biological differences between groups.Mutational landscapes and tumor immune microenvironments were compared.Sensitivity to immunotherapy and antineoplastic drugs was also predicted using PANI.The effects of Z-DNA-binding protein 1(ZBP1)on cell proliferation and migration were assessed by cell counting kit-8(CCK-8)and Transwell assays.Results We identified five PRGs(ZBP1,tumor necrosis factor superfamily protein 14(TNFSF14),cyclin-dependent kinase inhibitor 3(CDKN3),parathyroid hormone-like hormone(PTHLH),and heme-oxygenase 1(HMOX1))constituting PANI,independently associated with ccRCC patient prognosis.The PANI-based nomogram,integrated with clinical factors,demonstrated high predictive accuracy for prognosis.High-PANI patients exhibited distinct co-mutation patterns in ccRCC driver genes and lower survival probabilities,with an enriched immune-related functional profile,indicating an activated immune environment.These patients also showed increased sensitivity to immunotherapy and antineoplastic drugs.The knockdown of ZBP1,a key PRG in the PANI,significantly reduced ccRCC cell proliferation and migration.Conclusions PANI provides precise prognosis and immunotherapy response predictions for ccRCC patients,facilitating individualized treatment strategies.展开更多
Background:Clear cell renal carcinoma(ccRCC),the leading histological subtype of RCC,lacks any targeted therapy options.Although some studies have shown that early growth response factor 1(EGR1)has a significant role ...Background:Clear cell renal carcinoma(ccRCC),the leading histological subtype of RCC,lacks any targeted therapy options.Although some studies have shown that early growth response factor 1(EGR1)has a significant role in cancer development and progression,its role and underlying mechanisms in ccRCC remain poorly understood.Methods:The Cancer Genome Atlas(TCGA)database was utilized to examine the expression of EGR1 in ccRCC.The expression of EGR1 in 55 ccRCC tissues was evaluated using immunohistochemistry.The link between EGR1 expression and clinicopathological variables was examined through an analysis.Gain-of-function assays were employed to investigate EGR1’s biological functions in ccRCC cells,involving proliferation,colony formation,invasion assays,and tumorigenesis in nude mice.In order to assess the protein expression of mitogen-activated protein kinase 15(MAPK15),E-cadherin,matrix metalloproteinase-9/-2(MMP-9 and MMP-2),Western blot technique was applied.Results:The results revealed a decrease in EGR1 expression in ccRCC tissues.This decrease was strongly linked to TNM stage,lymphatic metastasis,tumor size,histological grade,and unfavorable prognosis in ccRCC patients.It has been demonstrated that overexpressing EGR1 inhibits the growth of xenograft tumors in vivo and inhibits cell colony formation,motility,and invasion in vitro.Furthermore,EGR1 can prevent the development and movement of ccRCC cells by controlling the expression of MMP-2,MMP-9,E-cadherin,and MAPK15.Conclusions:The EGR1/MAPK15 axis may represent a promising target for drug development,with EGR1 serving as a possible target for ccRCC therapy.展开更多
Background:Clear cell renal cell carcinoma(ccRCC)is an aggressive malignancy associated with limited treatment options and poor prognosis.Emerging studies suggest that the actin-regulating protein actin-related protei...Background:Clear cell renal cell carcinoma(ccRCC)is an aggressive malignancy associated with limited treatment options and poor prognosis.Emerging studies suggest that the actin-regulating protein actin-related protein 2/3 complex subunit 1B(ARPC1B),a key regulatory protein within the actin cytoskeleton,could play a pivotal role in ccRCC progression.The current study aimed to uncover the biological functions of ARPC1B and the molecular mechanisms driving its effects in ccRCC.Methods:ARPC1B expression and prognostic implications were analyzed using data sourced from the Gene Expression Profiling Interactive Analysis(GEPIA)platform,immunohistochemical(IHC)staining on 150 tumor samples along with 30 corresponding normal tissues,and Western blotting(WB)analyses across multiple ccRCC-derived cell lines.Functional assays assessing cell proliferation,colony formation capability,migration,invasion,and in vivo tumorigenicity were conducted following either ARPC1B suppression or upregulation.Additionally,WB analysis was utilized to evaluate proteins linked to epithelial-to-mesenchymal transition(EMT)and the Wnt/β-catenin pathway.Results:The findings revealed a substantial elevation of ARPC1B in ccRCC tissues and cell lines,significantly associated with advanced TNM stages,higher Fuhrman grades,and reduced overall survival(OS)(p<0.001).Multivariate statistical analysis identified ARPC1B as a standalone prognostic factor.Silencing ARPC1B notably impaired ccRCC cellular activities,and tumorigenesis in animalmodels,whereas augmented ARPC1B expression enhanced these malignant phenotypes.Mechanistically,downregulation of ARPC1B suppressed Wnt/β-catenin signaling and disrupted EMT,indicated by reducedβ-catenin,c-Myc,cyclin D1,and ZEB-1 levels,and concurrently increased E-cadherin expression.Additionally,reactivation of theWnt/β-catenin pathway partly reversed the inhibitory effects of ARPC1B depletion on tumor growth and invasiveness.Conclusions:ARPC1B emerges as an essential oncogenic factor in ccRCC by stimulating EMT and activating the Wnt/β-catenin pathway,ultimately enhancing tumor aggressiveness and metastatic potential.Thus,targeting ARPC1B represents a promising therapeutic strategy,warranting further exploration in ccRCC management.展开更多
基金supported by the Zhejiang Provincial Natural Science Foundation of China(No.LZ22H160008)the Zhejiang Medical Science and Technology Project(Nos.2022RC059 and 2023KY931),China.
文摘Objective To identify pyroptosis,apoptosis,and necroptosis(PANoptosis)-related genes(PRGs)in clear cell renal cell carcinoma(ccRCC)for patient stratification and prognosis prediction.Methods We used differential expression analysis and weighted gene co-expression network analysis(WGCNA)to identify ccRCC-specific PRGs.A prognostic model,the PANoptosis-index(PANI),was constructed using least absolute shrinkage and selection operator(LASSO)and Cox regression.The PANI model,comprising PRGs,was validated through single-cell RNA-sequencing(scRNA-seq),immunohistochemistry,and reverse transcription-quantitative polymerase chain reaction(RT-qPCR).Patient cohorts were categorized into high-and low-PANI groups,and the model’s performance was appraised using various metrics.External validation was performed with the E-MTAB-1980 dataset.Functional and gene set enrichment analyses distinguished biological differences between groups.Mutational landscapes and tumor immune microenvironments were compared.Sensitivity to immunotherapy and antineoplastic drugs was also predicted using PANI.The effects of Z-DNA-binding protein 1(ZBP1)on cell proliferation and migration were assessed by cell counting kit-8(CCK-8)and Transwell assays.Results We identified five PRGs(ZBP1,tumor necrosis factor superfamily protein 14(TNFSF14),cyclin-dependent kinase inhibitor 3(CDKN3),parathyroid hormone-like hormone(PTHLH),and heme-oxygenase 1(HMOX1))constituting PANI,independently associated with ccRCC patient prognosis.The PANI-based nomogram,integrated with clinical factors,demonstrated high predictive accuracy for prognosis.High-PANI patients exhibited distinct co-mutation patterns in ccRCC driver genes and lower survival probabilities,with an enriched immune-related functional profile,indicating an activated immune environment.These patients also showed increased sensitivity to immunotherapy and antineoplastic drugs.The knockdown of ZBP1,a key PRG in the PANI,significantly reduced ccRCC cell proliferation and migration.Conclusions PANI provides precise prognosis and immunotherapy response predictions for ccRCC patients,facilitating individualized treatment strategies.
基金supported by grants from the Shenzhen Basic Research Project(JCYJ20210324125803008)the Shenzhen Longhua District Science and Innovation Bureau Fund for Medical Institutions(2022143).
文摘Background:Clear cell renal carcinoma(ccRCC),the leading histological subtype of RCC,lacks any targeted therapy options.Although some studies have shown that early growth response factor 1(EGR1)has a significant role in cancer development and progression,its role and underlying mechanisms in ccRCC remain poorly understood.Methods:The Cancer Genome Atlas(TCGA)database was utilized to examine the expression of EGR1 in ccRCC.The expression of EGR1 in 55 ccRCC tissues was evaluated using immunohistochemistry.The link between EGR1 expression and clinicopathological variables was examined through an analysis.Gain-of-function assays were employed to investigate EGR1’s biological functions in ccRCC cells,involving proliferation,colony formation,invasion assays,and tumorigenesis in nude mice.In order to assess the protein expression of mitogen-activated protein kinase 15(MAPK15),E-cadherin,matrix metalloproteinase-9/-2(MMP-9 and MMP-2),Western blot technique was applied.Results:The results revealed a decrease in EGR1 expression in ccRCC tissues.This decrease was strongly linked to TNM stage,lymphatic metastasis,tumor size,histological grade,and unfavorable prognosis in ccRCC patients.It has been demonstrated that overexpressing EGR1 inhibits the growth of xenograft tumors in vivo and inhibits cell colony formation,motility,and invasion in vitro.Furthermore,EGR1 can prevent the development and movement of ccRCC cells by controlling the expression of MMP-2,MMP-9,E-cadherin,and MAPK15.Conclusions:The EGR1/MAPK15 axis may represent a promising target for drug development,with EGR1 serving as a possible target for ccRCC therapy.
文摘Background:Clear cell renal cell carcinoma(ccRCC)is an aggressive malignancy associated with limited treatment options and poor prognosis.Emerging studies suggest that the actin-regulating protein actin-related protein 2/3 complex subunit 1B(ARPC1B),a key regulatory protein within the actin cytoskeleton,could play a pivotal role in ccRCC progression.The current study aimed to uncover the biological functions of ARPC1B and the molecular mechanisms driving its effects in ccRCC.Methods:ARPC1B expression and prognostic implications were analyzed using data sourced from the Gene Expression Profiling Interactive Analysis(GEPIA)platform,immunohistochemical(IHC)staining on 150 tumor samples along with 30 corresponding normal tissues,and Western blotting(WB)analyses across multiple ccRCC-derived cell lines.Functional assays assessing cell proliferation,colony formation capability,migration,invasion,and in vivo tumorigenicity were conducted following either ARPC1B suppression or upregulation.Additionally,WB analysis was utilized to evaluate proteins linked to epithelial-to-mesenchymal transition(EMT)and the Wnt/β-catenin pathway.Results:The findings revealed a substantial elevation of ARPC1B in ccRCC tissues and cell lines,significantly associated with advanced TNM stages,higher Fuhrman grades,and reduced overall survival(OS)(p<0.001).Multivariate statistical analysis identified ARPC1B as a standalone prognostic factor.Silencing ARPC1B notably impaired ccRCC cellular activities,and tumorigenesis in animalmodels,whereas augmented ARPC1B expression enhanced these malignant phenotypes.Mechanistically,downregulation of ARPC1B suppressed Wnt/β-catenin signaling and disrupted EMT,indicated by reducedβ-catenin,c-Myc,cyclin D1,and ZEB-1 levels,and concurrently increased E-cadherin expression.Additionally,reactivation of theWnt/β-catenin pathway partly reversed the inhibitory effects of ARPC1B depletion on tumor growth and invasiveness.Conclusions:ARPC1B emerges as an essential oncogenic factor in ccRCC by stimulating EMT and activating the Wnt/β-catenin pathway,ultimately enhancing tumor aggressiveness and metastatic potential.Thus,targeting ARPC1B represents a promising therapeutic strategy,warranting further exploration in ccRCC management.
