Background:Numerous studies have revealed a tight connection between tumor development and the coagulation system.However,the effects of coagulation on the prognosis and tumor microenvironment(TME)of clear cell renal ...Background:Numerous studies have revealed a tight connection between tumor development and the coagulation system.However,the effects of coagulation on the prognosis and tumor microenvironment(TME)of clear cell renal cell carcinoma(ccRCC)remain poorly understood.Methods:We employed the consensus clustering method to characterize distinct molecular subtypes associated with coagulation patterns.Subsequently,we examined variations in the overall survival(OS),genomic profiles,and TME characteristics between these subtypes.To develop a prognostic coagulation-related risk score(CRRS)model,we utilized the least absolute shrinkage and selection operator Cox regression and stepwise multivariate Cox regression analyses.We also created a nomogram to aid in the clinical application of the risk score,evaluating the relationships between the CRRS and the immune microenvironment,responsiveness to immunotherapy,and targeted treatment.The clinical significance of PLAUR and its biological function in ccRCC were also further analyzed.Results:There were significant differences in clinical features,prognostic stratification,genomic variation,and TME characteristics between the two coagulation-related subtypes.We established and validated a CRRS using six coagulation-related genes that can be employed as an effective indicator of risk stratification and prognosis estimation for ccRCC patients.Significant variations in survival outcomes were observed between the high-and low-risk groups.The nomogram was proficient in predicting the 1-,3-,and 5-year OS.Additionally,the CRRS emerged as a novel tool for evaluating the clinical effectiveness of immunotherapy and targeted treatments in ccRCC.Moreover,we confirmed upregulated PLAUR expression in ccRCC samples that was significantly correlated with poor patient prognosis.PLAUR knockdown notably inhibited ccRCC cell proliferation and migration.Conclusion:Our data suggested that CRRS may be employed as a reliable predictive biomarker that can provide therapeutic benefits for immunotherapy and targeted therapy in ccRCC.展开更多
Embryonic stem （ES） cells are potent resources for cell therapy,and monoclonal antibodies （mAbs） against native cell surface markers of ES cells could be useful tools for therapeutic applications.Here,we report th...Embryonic stem （ES） cells are potent resources for cell therapy,and monoclonal antibodies （mAbs） against native cell surface markers of ES cells could be useful tools for therapeutic applications.Here,we report the development of a feasible approach,which could be used in mass production,for experimentally producing rabbit mAbs against native cell surface antigens on the cell surface.Two of the 14 mAbs,which were selected at random,could be bound to the cell surface antigens of mES cells.The immunocytochemistry （ICC） and Western blot results showed that mAb 39 recognises conformational epitopes.The target antigen of mAb 39 was then successfully purified using an improved immunoprecipitation approach in which mAb was bounded to intact mES cells before the cells were lysed.The LC-LTQ mass spectrum analysis showed that the target antigen of mAb 39 was Glut3.This result was further confirmed by Western blot using commercially available antibodies against Glut3.Further experiments showed that mAb 39 exhibited an antiproliferative effect on mES cells.We also found that Glut3 was differentially expressed among the mES cell population as detected by flow cytometry.展开更多
文摘Background:Numerous studies have revealed a tight connection between tumor development and the coagulation system.However,the effects of coagulation on the prognosis and tumor microenvironment(TME)of clear cell renal cell carcinoma(ccRCC)remain poorly understood.Methods:We employed the consensus clustering method to characterize distinct molecular subtypes associated with coagulation patterns.Subsequently,we examined variations in the overall survival(OS),genomic profiles,and TME characteristics between these subtypes.To develop a prognostic coagulation-related risk score(CRRS)model,we utilized the least absolute shrinkage and selection operator Cox regression and stepwise multivariate Cox regression analyses.We also created a nomogram to aid in the clinical application of the risk score,evaluating the relationships between the CRRS and the immune microenvironment,responsiveness to immunotherapy,and targeted treatment.The clinical significance of PLAUR and its biological function in ccRCC were also further analyzed.Results:There were significant differences in clinical features,prognostic stratification,genomic variation,and TME characteristics between the two coagulation-related subtypes.We established and validated a CRRS using six coagulation-related genes that can be employed as an effective indicator of risk stratification and prognosis estimation for ccRCC patients.Significant variations in survival outcomes were observed between the high-and low-risk groups.The nomogram was proficient in predicting the 1-,3-,and 5-year OS.Additionally,the CRRS emerged as a novel tool for evaluating the clinical effectiveness of immunotherapy and targeted treatments in ccRCC.Moreover,we confirmed upregulated PLAUR expression in ccRCC samples that was significantly correlated with poor patient prognosis.PLAUR knockdown notably inhibited ccRCC cell proliferation and migration.Conclusion:Our data suggested that CRRS may be employed as a reliable predictive biomarker that can provide therapeutic benefits for immunotherapy and targeted therapy in ccRCC.
基金supported by the National Key Scientific Research Program of China,the Ministry of Science and Technology of the People’s Republic of China (No 2007947804)
文摘Embryonic stem （ES） cells are potent resources for cell therapy,and monoclonal antibodies （mAbs） against native cell surface markers of ES cells could be useful tools for therapeutic applications.Here,we report the development of a feasible approach,which could be used in mass production,for experimentally producing rabbit mAbs against native cell surface antigens on the cell surface.Two of the 14 mAbs,which were selected at random,could be bound to the cell surface antigens of mES cells.The immunocytochemistry （ICC） and Western blot results showed that mAb 39 recognises conformational epitopes.The target antigen of mAb 39 was then successfully purified using an improved immunoprecipitation approach in which mAb was bounded to intact mES cells before the cells were lysed.The LC-LTQ mass spectrum analysis showed that the target antigen of mAb 39 was Glut3.This result was further confirmed by Western blot using commercially available antibodies against Glut3.Further experiments showed that mAb 39 exhibited an antiproliferative effect on mES cells.We also found that Glut3 was differentially expressed among the mES cell population as detected by flow cytometry.
