Objective Pancreatic cancer is one of the most deadly cancers, which is characterized by its high metastatic potential. S100A4 is a major prometastatic protein involved in tumor invasion and metastasis which precise r...Objective Pancreatic cancer is one of the most deadly cancers, which is characterized by its high metastatic potential. S100A4 is a major prometastatic protein involved in tumor invasion and metastasis which precise role in pancreatic cancer has not been fully investigated. We knocked down the S100A4 gene in the Bxpc-3 pancreatic cancer cell line via RNA interference to study the changes in cell behavior. Methods Real-time polymerase chain reaction and western blotting were used to detect mRNA and protein expression levels of S100A4, matrix metalloproteinase （MMP）-2, E-cadherin and thrombospondin （TSP）-I. Transwell chambers were used to detect the migration and invasion abilities; a cell adhesion assay was used to detect adhesion ability; colony forming efficiency was used to detect cell proliferation; flow cytometry was used to detect apoptosis. Results S100A4 mRNA expression was reduced to 17% after transfection with SIOOA4-siRNA, and protein expression had a similar trend, mRNA and protein expression of MMP-2 was reduced and that of E-cadherin and TSP-1 was elevated, indicating that S100A4 affects their expression. S100A4-silenced cells exhibited a marked decrease in migration and invasiveness and increased adhesion, whereas overall proliferation and apoptosis were not overtly altered. Conclusion S100A4 and its downstream factors play important roles in pancreatic cancer invasion, and silencing AIOOA4 can significantly contain the invasiveness of pancreatic cancer.展开更多
Previously, we demonstrated that macrophages from thrombospondin 1 （TSP1）-deficient mice have a reduced inflammatory phenotype, suggesting that TSP1 plays a role in macrophage activation. In this study, we determine...Previously, we demonstrated that macrophages from thrombospondin 1 （TSP1）-deficient mice have a reduced inflammatory phenotype, suggesting that TSP1 plays a role in macrophage activation. In this study, we determined how TSP1 regulates macrophage function. We found that recombinant or purified piatelet human TSP1 treatment stimulated tumor-necrosis factor （TNF）-α expression in bone marrow-derived macrophages in a time- and dose-dependent manner. Toll-like receptor 4 （TLR4） expression （at the mRNA and protein levels） and nuclear factor-kappaB （NF-KB） activity were also stimulated by TSP1 treatment. The TSPl-mediated increase in TNF-a production was abolished in TLR4-deficient macrophages, suggesting that TSP1 activates macrophages through a TLR4-dependent pathway. TSP1 also stimulated TLR4 activation in macrophages in vivo. Furthermore, TSPl-mediated macrophage activation was attenuated by using a peptide or an antibody to block the association between TSP1 and CD36. Taken together, these data suggest that the stimulation of the macrophage TLR4 pathway by TSP1 is partially mediated by the interaction of TSP1 with its receptor, CD36.展开更多
背景:目前对骨关节患者滑膜、关节软骨中相关降解酶的研究较多,对关节液中相关因子的检测亦有报道,但有关不同病变分期相关因子表达情况的报道较少,且其与病变程度相关性研究报道亦相对较少。目的:检测不同分期骨关节炎患者关节液中细...背景:目前对骨关节患者滑膜、关节软骨中相关降解酶的研究较多,对关节液中相关因子的检测亦有报道,但有关不同病变分期相关因子表达情况的报道较少,且其与病变程度相关性研究报道亦相对较少。目的:检测不同分期骨关节炎患者关节液中细胞外调节蛋白激酶、基质金属蛋白酶13和人类含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶4/5(a disintegrin and metalloproteinase with thrombospondin-like motifs,ADAMTS4/5)的表达。方法:选择2016年10月至2017年12月在石河子大学医学院第一附属医院骨科行关节镜手术的骨关节炎患者94例,根据临床症状与K-L X射线分级分为早期组(n=27)、中期组(n=32)、晚期组(n=35);选择行截肢的健康患者、单纯膝外伤行关节镜检查者及自愿参与试验的门诊体检者10例,作为对照组。采集4组受试者关节液,采用荧光定量PCR法和ELISA法检测细胞外调节蛋白激酶、基质金属蛋白酶13和ADAMTS4/5的表达情况。试验经石河子大学医学院第一附属医院医学伦理委员会批准,批准号:2017-052-01。结果与结论:①荧光定量PCR检测:早、中、晚期组的各基因表达均高于对照组(P<0.001),早、中、晚期组的细胞外调节蛋白激酶、基质金属蛋白酶13基因表达呈逐渐上升趋势(P<0.01),早期组ADAMTS4/5基因表达高于中、晚期组(P<0.01);②ELISA检测:早、中、晚期组的关节液中各因子表达均高于对照组(P<0.001),早、中、晚期组的细胞外调节蛋白激酶、基质金属蛋白酶13表达呈逐渐上升趋势(P<0.01),早期组ADAMTS4/5表达高于中、晚期组(P<0.01);③相关性分析:细胞外调节蛋白激酶与基质金属蛋白酶13表达水平呈正相关关系(P<0.01),ADAMTS4与ADAMTS5表达呈正相关关系(P<0.01),ADAMTS4、ADAMTS5与细胞外调节蛋白激酶、基质金属蛋白酶13表达水平呈负相关关系(P<0.05);④结果表明:细胞外调节蛋白激酶、基质金属蛋白�展开更多
文摘Objective Pancreatic cancer is one of the most deadly cancers, which is characterized by its high metastatic potential. S100A4 is a major prometastatic protein involved in tumor invasion and metastasis which precise role in pancreatic cancer has not been fully investigated. We knocked down the S100A4 gene in the Bxpc-3 pancreatic cancer cell line via RNA interference to study the changes in cell behavior. Methods Real-time polymerase chain reaction and western blotting were used to detect mRNA and protein expression levels of S100A4, matrix metalloproteinase （MMP）-2, E-cadherin and thrombospondin （TSP）-I. Transwell chambers were used to detect the migration and invasion abilities; a cell adhesion assay was used to detect adhesion ability; colony forming efficiency was used to detect cell proliferation; flow cytometry was used to detect apoptosis. Results S100A4 mRNA expression was reduced to 17% after transfection with SIOOA4-siRNA, and protein expression had a similar trend, mRNA and protein expression of MMP-2 was reduced and that of E-cadherin and TSP-1 was elevated, indicating that S100A4 affects their expression. S100A4-silenced cells exhibited a marked decrease in migration and invasiveness and increased adhesion, whereas overall proliferation and apoptosis were not overtly altered. Conclusion S100A4 and its downstream factors play important roles in pancreatic cancer invasion, and silencing AIOOA4 can significantly contain the invasiveness of pancreatic cancer.
文摘Previously, we demonstrated that macrophages from thrombospondin 1 （TSP1）-deficient mice have a reduced inflammatory phenotype, suggesting that TSP1 plays a role in macrophage activation. In this study, we determined how TSP1 regulates macrophage function. We found that recombinant or purified piatelet human TSP1 treatment stimulated tumor-necrosis factor （TNF）-α expression in bone marrow-derived macrophages in a time- and dose-dependent manner. Toll-like receptor 4 （TLR4） expression （at the mRNA and protein levels） and nuclear factor-kappaB （NF-KB） activity were also stimulated by TSP1 treatment. The TSPl-mediated increase in TNF-a production was abolished in TLR4-deficient macrophages, suggesting that TSP1 activates macrophages through a TLR4-dependent pathway. TSP1 also stimulated TLR4 activation in macrophages in vivo. Furthermore, TSPl-mediated macrophage activation was attenuated by using a peptide or an antibody to block the association between TSP1 and CD36. Taken together, these data suggest that the stimulation of the macrophage TLR4 pathway by TSP1 is partially mediated by the interaction of TSP1 with its receptor, CD36.
文摘背景:目前对骨关节患者滑膜、关节软骨中相关降解酶的研究较多,对关节液中相关因子的检测亦有报道,但有关不同病变分期相关因子表达情况的报道较少,且其与病变程度相关性研究报道亦相对较少。目的:检测不同分期骨关节炎患者关节液中细胞外调节蛋白激酶、基质金属蛋白酶13和人类含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶4/5(a disintegrin and metalloproteinase with thrombospondin-like motifs,ADAMTS4/5)的表达。方法:选择2016年10月至2017年12月在石河子大学医学院第一附属医院骨科行关节镜手术的骨关节炎患者94例,根据临床症状与K-L X射线分级分为早期组(n=27)、中期组(n=32)、晚期组(n=35);选择行截肢的健康患者、单纯膝外伤行关节镜检查者及自愿参与试验的门诊体检者10例,作为对照组。采集4组受试者关节液,采用荧光定量PCR法和ELISA法检测细胞外调节蛋白激酶、基质金属蛋白酶13和ADAMTS4/5的表达情况。试验经石河子大学医学院第一附属医院医学伦理委员会批准,批准号:2017-052-01。结果与结论:①荧光定量PCR检测:早、中、晚期组的各基因表达均高于对照组(P<0.001),早、中、晚期组的细胞外调节蛋白激酶、基质金属蛋白酶13基因表达呈逐渐上升趋势(P<0.01),早期组ADAMTS4/5基因表达高于中、晚期组(P<0.01);②ELISA检测:早、中、晚期组的关节液中各因子表达均高于对照组(P<0.001),早、中、晚期组的细胞外调节蛋白激酶、基质金属蛋白酶13表达呈逐渐上升趋势(P<0.01),早期组ADAMTS4/5表达高于中、晚期组(P<0.01);③相关性分析:细胞外调节蛋白激酶与基质金属蛋白酶13表达水平呈正相关关系(P<0.01),ADAMTS4与ADAMTS5表达呈正相关关系(P<0.01),ADAMTS4、ADAMTS5与细胞外调节蛋白激酶、基质金属蛋白酶13表达水平呈负相关关系(P<0.05);④结果表明:细胞外调节蛋白激酶、基质金属蛋白�
