目的本研究旨在探讨转录共激活因子(transcriptional coactivator with PDZ-binding motif,TAZ)与组蛋白H2A(histone H2A,H2A)的一种组蛋白变体H2A.Z,在缺氧诱导的肺损伤后修复中的作用及其潜在机制。方法利用缺氧模拟仓(5800 m)处理4 d...目的本研究旨在探讨转录共激活因子(transcriptional coactivator with PDZ-binding motif,TAZ)与组蛋白H2A(histone H2A,H2A)的一种组蛋白变体H2A.Z,在缺氧诱导的肺损伤后修复中的作用及其潜在机制。方法利用缺氧模拟仓(5800 m)处理4 d后,构建小鼠缺氧肺损伤模型,肺组织切片HE染色评估肺损伤程度。利用缺氧工作站(1%O_(2)浓度)处理小鼠肺泡上皮细胞系(murine lung epithelial-12,MLE12)24 h,构建MLE12缺氧细胞模型,Western blot分析检测TAZ表达。CCK-8实验评价肺泡上皮细胞(alveolar epithelial cells,AECs)增殖。免疫共沉淀(co-immunoprecipitation,Co-IP)实验验证TAZ与H2A.Z二者之间的相互作用。靶向剪切及转座酶标记技术(cleavage under targets and tagmentation,CUT&Tag)测序分析TAZ影响H2A.Z在染色质沉积情况。结果缺氧显著诱导小鼠肺组织肺泡萎缩及炎症浸润(P<0.01)。缺氧培养显著上调MLE12细胞中TAZ蛋白表达(P<0.05)。CCK-8实验发现,敲低TAZ后AECs增殖能力显著下降(P<0.01)。Co-IP证实TAZ与H2A.Z存在物理相互作用。靶向剪切及转座酶标记技术(cleavage under targets and tagmentation,CUT&Tag)测序显示缺氧促进H2A.Z在染色质上的沉积(常氧结合位点31817,缺氧结合位点44078),而TAZ敲低部分逆转此现象(结合位点37840)。结论缺氧显著上调TAZ表达,后者与H2A.Z的结合并促进H2A.Z在染色质上的沉积,增强AECs增殖以应对缺氧损伤。展开更多
目的:研究食管癌及癌旁组织中PDZ结合域的转录共刺激因子(transcriptional co-activator with PDZ-binding motif,TAZ)蛋白的表达,并探讨其临床意义。方法:采用免疫组化SP法检测123例食管癌及对应癌旁组织中TAZ蛋白的表达,并对其与临床...目的:研究食管癌及癌旁组织中PDZ结合域的转录共刺激因子(transcriptional co-activator with PDZ-binding motif,TAZ)蛋白的表达,并探讨其临床意义。方法:采用免疫组化SP法检测123例食管癌及对应癌旁组织中TAZ蛋白的表达,并对其与临床特征的关系作相关统计分析。结果:TAZ蛋白在食管癌组织中的阳性表达率明显高于对应癌旁组织(60.2%vs.18.7%,P<0.001),且其阳性表达与患者临床分期晚、肿瘤分化程度低、总生存期短相关(P均<0.05);COX比例多因素风险模型分析显示,TAZ阳性表达是食管癌预后的独立影响因子(P=0.031)。结论:TAZ蛋白在食管癌组织中表达升高,是食管癌预后的独立影响因子,TAZ蛋白有望成为食管癌诊断和预后判断的一个新指标。展开更多
目的探讨溶血磷脂酸(LPA)、Yes相关蛋白(YAP)及含PDZ结合区的共转录激活因子(TAZ)在头颈鳞癌中的表达及其与临床病理特征的关系。方法选择性收集雅安市人民医院病理科2010年1月至2016年12月期间,手术切除的病理组织石蜡标本共计136例,...目的探讨溶血磷脂酸(LPA)、Yes相关蛋白(YAP)及含PDZ结合区的共转录激活因子(TAZ)在头颈鳞癌中的表达及其与临床病理特征的关系。方法选择性收集雅安市人民医院病理科2010年1月至2016年12月期间,手术切除的病理组织石蜡标本共计136例,其中包括头颈鳞癌癌旁正常组织37例、头颈良性瘤样组织35例、头颈鳞癌64例,所有标本术前均无放化疗史,采用免疫组化及实时定量PCR检测LPA、YAP及TAZ在各组织中的表达及临床相关参数的关系。结果 LPA m RNA在头颈鳞癌癌旁正常组织、良性瘤样组织和鳞癌组织中的表达相近(P>0.05);YAP m RNA在鳞癌组织中的表达要高于癌旁组织、瘤样组织中的表达(P<0.05);TAZ m RNA在头颈3种组织中呈低表达,但鳞癌组织的表达明显高于良性瘤样组织及癌旁正常组织(P<0.05);YAP及TAZ蛋白在头颈鳞癌组织中的表达显著高于在癌旁正常组织和良性瘤样组织中的表达(P<0.05);鳞癌患者YAP的表达在肿瘤>3.0 cm、淋巴结阳性显著高于对照组(P<0.05);TAZ的表达在肿瘤中低分化、肿瘤>3.0 cm、淋巴结阳性及Ⅲ~Ⅳ期高于对照组(P<0.05)。结论 YAP及TAZ在头颈鳞癌中的表达与肿瘤分期、分化程度、淋巴结转移密切相关,而LPA的表达无显著差异及相关性。展开更多
It is critical to combat tumor metastasis by eradicating disseminated tumor cells in any step during the metastasis process.After entering the blood circulation system,tumor cells are in suspension and experience cons...It is critical to combat tumor metastasis by eradicating disseminated tumor cells in any step during the metastasis process.After entering the blood circulation system,tumor cells are in suspension and experience considerable levels of fluid shear stress.However,the influence of hemodynamic shear stress on the survival of CTCs and the underlying mechanotransduction mechanisms remain unclear.This study shows that fluid shear stress can eliminate the majority of CTCs and the viability of suspended tumor cells depends on the stress magnitude,indicating that tumor cells can sense and respond to fluid shear stress.Mechanistically,the expression of Piezo1 but not Piezo2 is greatly upregulated in suspended tumor cells after shear stress treatment.Inhibiting/activating Piezo1 increases/decreases the viability of suspended tumor cells in shear flow,which depends on Piezo1-mediated calcium entry.These findings suggest that Piezo1 may be the major mechanosensor by which suspended tumor cells sense fluid shear stress.As the downstream effector of Piezo1,actomyosin in tumor cells is significantly activated under increasing shear stress.Its activity influences the survival of CTCs in shear flow and rescues the effects of Piezo1 on tumor cell survival,suggesting that hemodynamic shear stress regulates the survival of CTCs through Piezo1 mediated actomyosin activity.Importantly,fluid shear stress considerably up-regulates YAP/TAZ activity of suspended tumor cells and promotes their nuclear translocation in a magnitude-dependent manner.Inhibiting YAP/TAZ enhances the viability of suspended tumor cells in shear stress,while activating their activity decreases tumor cell survival,suggesting that YAP/TAZ activation promotes the apoptosis of suspended tumor cells,which is different from the findings that YAP/TAZ facilitates the survival of adherent cells to shear flow.Further,blocking the nuclear import of YAP/TAZ inactivates the sensation of suspended tumor cells to fluid shear flow and attenuates the dependence of tumor cell survival on different magnitudes of hemodynamic shear stress.The influence of Piezo1-actomyosin pathway on suspended tumor cells can be rescued by YAP/TAZ activity,suggesting that Piezo1-mediated signaling induces tumor cell apoptosis via nuclear translocation of YAP/TAZ.In addition,fluid shear stress can also activate the expressions of LATS1/2 and MST1/2 in Hippo pathway through Piezo1,which is known to inhibit YAP/TAZ activity.Silencing/activating LATS1/2 or MST1/2 inhibits/enhances the viability of CTCs under shear stress,the effects of which can be further rescued by YAP/TAZ.These findings suggest that the responses of suspended tumor cells to hemodynamic shear stress are partially mediated by Hippo signaling.After nuclear translocation,YAP/TAZ directly bind p73/PUMA,which further promotes the transcription of pro-apoptotic genes and induces the apoptosis of suspended tumor cells.In summary,these findings show that hemodynamic shear stress considerably influences the survival of CTCs in blood circulation.We have identified the calcium channel Piezo1 as a novel mechanosensor for the response of CTCs to fluid shear stress.Hemodynamic shear stress induces the apoptosis of suspended tumor cells through Piezo1-actomyosin-YAP/TAZ-p73/PUMA signaling,which is different from the mechanotranduction mechanisms for tumor cells in adherent.Therefore,this study has unveiled the novel mechanosensor of suspended CTCs in response to fluid shear stress and the subsequent mechanisms and identified Piezo1 and YAP/TAZ as the potential therapeutic targets,through which CTCs may be effectively eradicated in the vasculature to prohibit tumor metastasis.展开更多
文摘目的本研究旨在探讨转录共激活因子(transcriptional coactivator with PDZ-binding motif,TAZ)与组蛋白H2A(histone H2A,H2A)的一种组蛋白变体H2A.Z,在缺氧诱导的肺损伤后修复中的作用及其潜在机制。方法利用缺氧模拟仓(5800 m)处理4 d后,构建小鼠缺氧肺损伤模型,肺组织切片HE染色评估肺损伤程度。利用缺氧工作站(1%O_(2)浓度)处理小鼠肺泡上皮细胞系(murine lung epithelial-12,MLE12)24 h,构建MLE12缺氧细胞模型,Western blot分析检测TAZ表达。CCK-8实验评价肺泡上皮细胞(alveolar epithelial cells,AECs)增殖。免疫共沉淀(co-immunoprecipitation,Co-IP)实验验证TAZ与H2A.Z二者之间的相互作用。靶向剪切及转座酶标记技术(cleavage under targets and tagmentation,CUT&Tag)测序分析TAZ影响H2A.Z在染色质沉积情况。结果缺氧显著诱导小鼠肺组织肺泡萎缩及炎症浸润(P<0.01)。缺氧培养显著上调MLE12细胞中TAZ蛋白表达(P<0.05)。CCK-8实验发现,敲低TAZ后AECs增殖能力显著下降(P<0.01)。Co-IP证实TAZ与H2A.Z存在物理相互作用。靶向剪切及转座酶标记技术(cleavage under targets and tagmentation,CUT&Tag)测序显示缺氧促进H2A.Z在染色质上的沉积(常氧结合位点31817,缺氧结合位点44078),而TAZ敲低部分逆转此现象(结合位点37840)。结论缺氧显著上调TAZ表达,后者与H2A.Z的结合并促进H2A.Z在染色质上的沉积,增强AECs增殖以应对缺氧损伤。
文摘目的:研究食管癌及癌旁组织中PDZ结合域的转录共刺激因子(transcriptional co-activator with PDZ-binding motif,TAZ)蛋白的表达,并探讨其临床意义。方法:采用免疫组化SP法检测123例食管癌及对应癌旁组织中TAZ蛋白的表达,并对其与临床特征的关系作相关统计分析。结果:TAZ蛋白在食管癌组织中的阳性表达率明显高于对应癌旁组织(60.2%vs.18.7%,P<0.001),且其阳性表达与患者临床分期晚、肿瘤分化程度低、总生存期短相关(P均<0.05);COX比例多因素风险模型分析显示,TAZ阳性表达是食管癌预后的独立影响因子(P=0.031)。结论:TAZ蛋白在食管癌组织中表达升高,是食管癌预后的独立影响因子,TAZ蛋白有望成为食管癌诊断和预后判断的一个新指标。
文摘目的探讨溶血磷脂酸(LPA)、Yes相关蛋白(YAP)及含PDZ结合区的共转录激活因子(TAZ)在头颈鳞癌中的表达及其与临床病理特征的关系。方法选择性收集雅安市人民医院病理科2010年1月至2016年12月期间,手术切除的病理组织石蜡标本共计136例,其中包括头颈鳞癌癌旁正常组织37例、头颈良性瘤样组织35例、头颈鳞癌64例,所有标本术前均无放化疗史,采用免疫组化及实时定量PCR检测LPA、YAP及TAZ在各组织中的表达及临床相关参数的关系。结果 LPA m RNA在头颈鳞癌癌旁正常组织、良性瘤样组织和鳞癌组织中的表达相近(P>0.05);YAP m RNA在鳞癌组织中的表达要高于癌旁组织、瘤样组织中的表达(P<0.05);TAZ m RNA在头颈3种组织中呈低表达,但鳞癌组织的表达明显高于良性瘤样组织及癌旁正常组织(P<0.05);YAP及TAZ蛋白在头颈鳞癌组织中的表达显著高于在癌旁正常组织和良性瘤样组织中的表达(P<0.05);鳞癌患者YAP的表达在肿瘤>3.0 cm、淋巴结阳性显著高于对照组(P<0.05);TAZ的表达在肿瘤中低分化、肿瘤>3.0 cm、淋巴结阳性及Ⅲ~Ⅳ期高于对照组(P<0.05)。结论 YAP及TAZ在头颈鳞癌中的表达与肿瘤分期、分化程度、淋巴结转移密切相关,而LPA的表达无显著差异及相关性。
文摘It is critical to combat tumor metastasis by eradicating disseminated tumor cells in any step during the metastasis process.After entering the blood circulation system,tumor cells are in suspension and experience considerable levels of fluid shear stress.However,the influence of hemodynamic shear stress on the survival of CTCs and the underlying mechanotransduction mechanisms remain unclear.This study shows that fluid shear stress can eliminate the majority of CTCs and the viability of suspended tumor cells depends on the stress magnitude,indicating that tumor cells can sense and respond to fluid shear stress.Mechanistically,the expression of Piezo1 but not Piezo2 is greatly upregulated in suspended tumor cells after shear stress treatment.Inhibiting/activating Piezo1 increases/decreases the viability of suspended tumor cells in shear flow,which depends on Piezo1-mediated calcium entry.These findings suggest that Piezo1 may be the major mechanosensor by which suspended tumor cells sense fluid shear stress.As the downstream effector of Piezo1,actomyosin in tumor cells is significantly activated under increasing shear stress.Its activity influences the survival of CTCs in shear flow and rescues the effects of Piezo1 on tumor cell survival,suggesting that hemodynamic shear stress regulates the survival of CTCs through Piezo1 mediated actomyosin activity.Importantly,fluid shear stress considerably up-regulates YAP/TAZ activity of suspended tumor cells and promotes their nuclear translocation in a magnitude-dependent manner.Inhibiting YAP/TAZ enhances the viability of suspended tumor cells in shear stress,while activating their activity decreases tumor cell survival,suggesting that YAP/TAZ activation promotes the apoptosis of suspended tumor cells,which is different from the findings that YAP/TAZ facilitates the survival of adherent cells to shear flow.Further,blocking the nuclear import of YAP/TAZ inactivates the sensation of suspended tumor cells to fluid shear flow and attenuates the dependence of tumor cell survival on different magnitudes of hemodynamic shear stress.The influence of Piezo1-actomyosin pathway on suspended tumor cells can be rescued by YAP/TAZ activity,suggesting that Piezo1-mediated signaling induces tumor cell apoptosis via nuclear translocation of YAP/TAZ.In addition,fluid shear stress can also activate the expressions of LATS1/2 and MST1/2 in Hippo pathway through Piezo1,which is known to inhibit YAP/TAZ activity.Silencing/activating LATS1/2 or MST1/2 inhibits/enhances the viability of CTCs under shear stress,the effects of which can be further rescued by YAP/TAZ.These findings suggest that the responses of suspended tumor cells to hemodynamic shear stress are partially mediated by Hippo signaling.After nuclear translocation,YAP/TAZ directly bind p73/PUMA,which further promotes the transcription of pro-apoptotic genes and induces the apoptosis of suspended tumor cells.In summary,these findings show that hemodynamic shear stress considerably influences the survival of CTCs in blood circulation.We have identified the calcium channel Piezo1 as a novel mechanosensor for the response of CTCs to fluid shear stress.Hemodynamic shear stress induces the apoptosis of suspended tumor cells through Piezo1-actomyosin-YAP/TAZ-p73/PUMA signaling,which is different from the mechanotranduction mechanisms for tumor cells in adherent.Therefore,this study has unveiled the novel mechanosensor of suspended CTCs in response to fluid shear stress and the subsequent mechanisms and identified Piezo1 and YAP/TAZ as the potential therapeutic targets,through which CTCs may be effectively eradicated in the vasculature to prohibit tumor metastasis.
