Integrins are a family of transmembrane glycoproteins that mediate cell-cell and cell-extracellular matrix interactions. The integrin α4 subunit is widely expressed by cells from the immune system and its expression ...Integrins are a family of transmembrane glycoproteins that mediate cell-cell and cell-extracellular matrix interactions. The integrin α4 subunit is widely expressed by cells from the immune system and its expression by non-hematopoietic cells is scarce. In the present study, gene and protein expression of this integrin subunit was characterized in proliferating and quiescent human RPE cells. Immunofluorescent studies confirm that the α4 subunit is expressed in vitro by RPE cells, a result that has been validated by immunofluorescence and FACS analyses. The accumulation of the α4 integrin at cell-cell junctions in post-confluent RPE cell cultures negatively correlated with the level of expression of the mRNA transcript. Accordingly, transient transfection analyses reveal that the α4 promoter activity is considerably reduced when RPE cells form a confluent monolayer. Moreover, transfection of recombinant constructs bearing 5’-deletions of the α4 promoter segment allows the localization of strong negative regulatory elements on the -76 to -300 region of the α4 gene suggesting that its expression is intimately linked to the proliferative state of primary cultured RPE cells.展开更多
目的探讨β4整合素结合蛋白ITGB4BP对Wnt/β-catenin信号通路的影响。方法将对照组空载体pCMV-3B质粒和实验组pCMV-ITGB4BP质粒分别转染至HEK293细胞中,用Western blot和激光共聚焦技术检测细胞中β-catenin含量的变化,用含TCF/LEF启动...目的探讨β4整合素结合蛋白ITGB4BP对Wnt/β-catenin信号通路的影响。方法将对照组空载体pCMV-3B质粒和实验组pCMV-ITGB4BP质粒分别转染至HEK293细胞中,用Western blot和激光共聚焦技术检测细胞中β-catenin含量的变化,用含TCF/LEF启动子的荧光素酶TOPFlash质粒和内参pRL-TK质粒共转染对照组和实验组的HEK293细胞,用荧光素酶活性分析法检测ITGB4BP转染后HEK293细胞TCF/LEF活性的变化。设不转染质粒组为正常组,对照组和实验组均分别设未加LiCl组和加LiCl组。结果 Western blot检测结果:与正常组或对照组比较,实验组细胞β-catenin表达显著减少(P<0.05)。加LiCl之后,β-catenin的表达显著增强(P<0.05),与未加有LiCl的正常组或对照组比较,加LiCl的实验组细胞内β-catenin仍有较高的表达,但差异无显著性。激光共聚焦技术观察:实验组β-catenin(绿色荧光)表达较对照组降低。加LiCl之后,β-catenin(绿色荧光)表达增强,与未加LiCl的对照组相比,加LiCl的实验组仍有较强的荧光强度。荧光素酶活性分析法检测:实验组的荧光素酶活性较对照组显著降低(P<0.05)。LiCl作用之后,Wnt/β-catenin信号通路活性明显增强(P<0.05),与未加LiCl的对照组相比,加LiCl的实验组仍有较强的荧光素酶活性(P<0.05)。结论ITGB4BP可以下调Wnt/β-catenin信号通路的活性。展开更多
Vedolizumab is a humanized monoclonal antibody and one of the safest biologics for the treatment ofboth forms of inflammatory bowel disease(IBD)-Crohn’s disease and ulcerative colitis.It targets theα4β7 integrinand...Vedolizumab is a humanized monoclonal antibody and one of the safest biologics for the treatment ofboth forms of inflammatory bowel disease(IBD)-Crohn’s disease and ulcerative colitis.It targets theα4β7 integrinand blocks leukocyte trafficking to the gut.Regardless of its efficacy in many patients,non-response to vedolizumabtreatment poses a significant clinical challenge.In this review,we synthesize recent findings on genomic,transcriptomic,proteomic,and cellular biomarkers of vedolizumab response,emphasizing their roles in predicting therapeutic outcomesand understanding non-responsiveness.Key insights include the identification of epigenetic and transcriptomicsignatures,the involvement of Th17 and IL-6 signaling,and the role of baseline inflammatory markers like albumin.Discrepancies in findings highlight the complexity of biomarker discovery and underscore the need for standardized,multiparametric approaches to refine personalized treatment strategies.By bridging knowledge gaps in vedolizumabresponsiveness,this review aims to advance biomarker-driven decision-making and improve outcomes for patientswith IBD.展开更多
文摘Integrins are a family of transmembrane glycoproteins that mediate cell-cell and cell-extracellular matrix interactions. The integrin α4 subunit is widely expressed by cells from the immune system and its expression by non-hematopoietic cells is scarce. In the present study, gene and protein expression of this integrin subunit was characterized in proliferating and quiescent human RPE cells. Immunofluorescent studies confirm that the α4 subunit is expressed in vitro by RPE cells, a result that has been validated by immunofluorescence and FACS analyses. The accumulation of the α4 integrin at cell-cell junctions in post-confluent RPE cell cultures negatively correlated with the level of expression of the mRNA transcript. Accordingly, transient transfection analyses reveal that the α4 promoter activity is considerably reduced when RPE cells form a confluent monolayer. Moreover, transfection of recombinant constructs bearing 5’-deletions of the α4 promoter segment allows the localization of strong negative regulatory elements on the -76 to -300 region of the α4 gene suggesting that its expression is intimately linked to the proliferative state of primary cultured RPE cells.
文摘目的探讨β4整合素结合蛋白ITGB4BP对Wnt/β-catenin信号通路的影响。方法将对照组空载体pCMV-3B质粒和实验组pCMV-ITGB4BP质粒分别转染至HEK293细胞中,用Western blot和激光共聚焦技术检测细胞中β-catenin含量的变化,用含TCF/LEF启动子的荧光素酶TOPFlash质粒和内参pRL-TK质粒共转染对照组和实验组的HEK293细胞,用荧光素酶活性分析法检测ITGB4BP转染后HEK293细胞TCF/LEF活性的变化。设不转染质粒组为正常组,对照组和实验组均分别设未加LiCl组和加LiCl组。结果 Western blot检测结果:与正常组或对照组比较,实验组细胞β-catenin表达显著减少(P<0.05)。加LiCl之后,β-catenin的表达显著增强(P<0.05),与未加有LiCl的正常组或对照组比较,加LiCl的实验组细胞内β-catenin仍有较高的表达,但差异无显著性。激光共聚焦技术观察:实验组β-catenin(绿色荧光)表达较对照组降低。加LiCl之后,β-catenin(绿色荧光)表达增强,与未加LiCl的对照组相比,加LiCl的实验组仍有较强的荧光强度。荧光素酶活性分析法检测:实验组的荧光素酶活性较对照组显著降低(P<0.05)。LiCl作用之后,Wnt/β-catenin信号通路活性明显增强(P<0.05),与未加LiCl的对照组相比,加LiCl的实验组仍有较强的荧光素酶活性(P<0.05)。结论ITGB4BP可以下调Wnt/β-catenin信号通路的活性。
基金supported by the Slovenian Research and Innovation Agency(ARIS)—Young Researcher Program(contract no.104-04/TK–6811)Research Core Funding P3-0427.
文摘Vedolizumab is a humanized monoclonal antibody and one of the safest biologics for the treatment ofboth forms of inflammatory bowel disease(IBD)-Crohn’s disease and ulcerative colitis.It targets theα4β7 integrinand blocks leukocyte trafficking to the gut.Regardless of its efficacy in many patients,non-response to vedolizumabtreatment poses a significant clinical challenge.In this review,we synthesize recent findings on genomic,transcriptomic,proteomic,and cellular biomarkers of vedolizumab response,emphasizing their roles in predicting therapeutic outcomesand understanding non-responsiveness.Key insights include the identification of epigenetic and transcriptomicsignatures,the involvement of Th17 and IL-6 signaling,and the role of baseline inflammatory markers like albumin.Discrepancies in findings highlight the complexity of biomarker discovery and underscore the need for standardized,multiparametric approaches to refine personalized treatment strategies.By bridging knowledge gaps in vedolizumabresponsiveness,this review aims to advance biomarker-driven decision-making and improve outcomes for patientswith IBD.
