BACKGROUND Yes-associated protein 1(YAP1),a downstream transcriptional coactivator regulated by the Hippo signaling pathway,has been shown to be involved in liver fibrosis.YAP activity is modulated by G-protein couple...BACKGROUND Yes-associated protein 1(YAP1),a downstream transcriptional coactivator regulated by the Hippo signaling pathway,has been shown to be involved in liver fibrosis.YAP activity is modulated by G-protein coupled receptors,including Gαs-coupled protein dopamine receptor D1(DRD1).Levodopa,a dopamine precursor,activates DRD1 on cell surface,triggering its downstream signaling pathway.AIM To investigate the therapeutic effect of levodopa and the downstream mechanism on carbon tetrachloride(CCl_(4))-induced liver fibrosis,including liver DRD1 expression.METHODS SD rats were intraperitoneally injected with 40%CCl_(4)for 8 weeks to induce liver fibrosis,followed by treatment with varying doses of levodopa for 2 weeks.Serum aspartate aminotransferase(AST)and alanine aminotransferase(ALT)levels were measured,and liver pathology was assessed using hematoxylin and eosin and Masson's staining.Alpha-smooth muscle actin(α-SMA)content,along with the expressions of DRD1,YAP,and phosphorylated protein,was analyzed by Western blot,immunohistochemistry,and reverse transcription-quantitative real-time polymerase chain reaction.RESULTS Compared with the controls,levodopa-treated rats showed a decrease in the proportion of collagen in the liver and a recovery from liver fibrosis(P=0.0007).Western blot and immunohistochemistry indicated that DRD1 was upregulated in the fibrotic liver of rats treated with levodopa,showing an increase in DRD1 Level(P<0.0001).In addition,the upregulation of DRD1 activated the Hippo signaling pathway,manifested as increased YAP phosphorylation(P<0.05).CONCLUSION This was the first study to demonstrate that levodopa attenuates CCl_(4)-induced liver fibrosis by inhibiting the Hippo/YAP signaling pathways.展开更多
目的:检测Hippo-YAP信号通路中主要效应蛋白Yes相关蛋白(Yes-associated protein,YAP)在正常胰腺组织和胰腺癌组织中的表达,探讨其表达强度与临床病理因素之间的关系及意义.方法:应用免疫组织化学EnVision法测定Hippo信号通路中YAP蛋白...目的:检测Hippo-YAP信号通路中主要效应蛋白Yes相关蛋白(Yes-associated protein,YAP)在正常胰腺组织和胰腺癌组织中的表达,探讨其表达强度与临床病理因素之间的关系及意义.方法:应用免疫组织化学EnVision法测定Hippo信号通路中YAP蛋白在45例胰腺癌及15例正常胰腺组织中的表达,应用RT-PCR方法检测YAP mRNA在45例胰腺癌及15例正常胰腺组织中的表达.结果:胰腺癌组织中YAP蛋白主要表达于胰腺导管上皮细胞,45例胰腺癌组织中YAP的阳性表达率为93.33%(42/45),15例正常胰腺组织中YAP阳性表达率为26.67%(4/15),两组间的表达差异具有统计学意义(2=27.95,P<0.01).在胰腺癌组织中YAP的表达与肿瘤分化程度密切相关(P=0.048),与年龄、性别、吸烟、饮酒、肥胖、糖耐量异常、糖尿病、慢性胰腺炎及临床分期等无关(P值分别是0.577、0.565、0.541、0.224、1.000、0.542、0.555、0.571、0.926).胰腺癌患者血清CA19-9水平与胰腺癌组织中YAP阳性程度呈正相关.YAP m R N A在胰腺癌组织中的表达较正常胰腺组织明显升高(0.3682±0.0783 vs 0.0394±0.0091,P<0.01).结论:Hippo-YAP通路可能在胰腺癌的发生、发展中发挥重要作用.并有可能成为胰腺癌治疗中新的靶点.展开更多
目的三阴性乳腺癌作为乳腺癌的特殊类型,尚缺乏有效的靶向治疗手段。文中旨在观察溶血磷脂酸(LPA)对三阴性乳腺癌的影响,探讨LPA-Hippo-Yes相关蛋白(YAP)信号通路在促进三阴性乳腺癌侵润转移中的作用及机制。方法体外合成YAP序列特异性...目的三阴性乳腺癌作为乳腺癌的特殊类型,尚缺乏有效的靶向治疗手段。文中旨在观察溶血磷脂酸(LPA)对三阴性乳腺癌的影响,探讨LPA-Hippo-Yes相关蛋白(YAP)信号通路在促进三阴性乳腺癌侵润转移中的作用及机制。方法体外合成YAP序列特异性小干扰RNA,应用质粒转染MDA-MB-231乳腺癌细胞株为实验组,非特异性siRNA转染MDA-MB-231细胞为阳性对照组,未转染MDA-MB-231细胞为空白对照组,每组设2个平行孔。检测各组对Hippo-YAP信号的影响,其作用机制及对Hippo-YAP上、下游信号通路调控。结果实验组总蛋白中YAP含量、细胞转染48h后的侵袭、转移能力[(0.035±0.005)、(2.200±1.000)个、(3.500±0.800)个]较阳性对照组[(0.343±0.012)、(27.600±5.100)个、(22.300±5.000)个]、空白对照组[(0.384±0.017)、(26.500±4.800)个、(22.350±6.000)个]明显降低(P<0.05)。实验组60、120、240 min时YAP表达量较阳性对照组、空白对照组显著降低(P<0.05)。实验组10、20、50μmol/L的YAP相对表达量显著低于阳性对照组、空白对照组(P<0.05)。C3转移酶和Y27632分别干预后,实验组p YAP mRNA含量(0.255±0.052、0.326±0.017)与空白对照组((0.048±0.032、0.534±0.017)、阳性对照组(0.052±0.021、0.528±0.024)比较差异有统计学意义(P<0.05)。实验组YAP m NA、AREG m NA的表达量(0.176±0.032、0.263±0.008)较空白对照组(0.043±0.013、0.051±0.014)、阴性对照组(0.049±0.025、0.057±0.043)显著升高(P<0.05)。结论 LPA诱导乳腺癌细胞侵袭转移,具有YAP蛋白依赖性,并具有时间与浓度依赖的特点;LPA-Hippo-YAP信号传导通路可能是促进三阴性乳腺癌细胞长时间转移的机制之一。展开更多
文摘BACKGROUND Yes-associated protein 1(YAP1),a downstream transcriptional coactivator regulated by the Hippo signaling pathway,has been shown to be involved in liver fibrosis.YAP activity is modulated by G-protein coupled receptors,including Gαs-coupled protein dopamine receptor D1(DRD1).Levodopa,a dopamine precursor,activates DRD1 on cell surface,triggering its downstream signaling pathway.AIM To investigate the therapeutic effect of levodopa and the downstream mechanism on carbon tetrachloride(CCl_(4))-induced liver fibrosis,including liver DRD1 expression.METHODS SD rats were intraperitoneally injected with 40%CCl_(4)for 8 weeks to induce liver fibrosis,followed by treatment with varying doses of levodopa for 2 weeks.Serum aspartate aminotransferase(AST)and alanine aminotransferase(ALT)levels were measured,and liver pathology was assessed using hematoxylin and eosin and Masson's staining.Alpha-smooth muscle actin(α-SMA)content,along with the expressions of DRD1,YAP,and phosphorylated protein,was analyzed by Western blot,immunohistochemistry,and reverse transcription-quantitative real-time polymerase chain reaction.RESULTS Compared with the controls,levodopa-treated rats showed a decrease in the proportion of collagen in the liver and a recovery from liver fibrosis(P=0.0007).Western blot and immunohistochemistry indicated that DRD1 was upregulated in the fibrotic liver of rats treated with levodopa,showing an increase in DRD1 Level(P<0.0001).In addition,the upregulation of DRD1 activated the Hippo signaling pathway,manifested as increased YAP phosphorylation(P<0.05).CONCLUSION This was the first study to demonstrate that levodopa attenuates CCl_(4)-induced liver fibrosis by inhibiting the Hippo/YAP signaling pathways.
文摘目的:检测Hippo-YAP信号通路中主要效应蛋白Yes相关蛋白(Yes-associated protein,YAP)在正常胰腺组织和胰腺癌组织中的表达,探讨其表达强度与临床病理因素之间的关系及意义.方法:应用免疫组织化学EnVision法测定Hippo信号通路中YAP蛋白在45例胰腺癌及15例正常胰腺组织中的表达,应用RT-PCR方法检测YAP mRNA在45例胰腺癌及15例正常胰腺组织中的表达.结果:胰腺癌组织中YAP蛋白主要表达于胰腺导管上皮细胞,45例胰腺癌组织中YAP的阳性表达率为93.33%(42/45),15例正常胰腺组织中YAP阳性表达率为26.67%(4/15),两组间的表达差异具有统计学意义(2=27.95,P<0.01).在胰腺癌组织中YAP的表达与肿瘤分化程度密切相关(P=0.048),与年龄、性别、吸烟、饮酒、肥胖、糖耐量异常、糖尿病、慢性胰腺炎及临床分期等无关(P值分别是0.577、0.565、0.541、0.224、1.000、0.542、0.555、0.571、0.926).胰腺癌患者血清CA19-9水平与胰腺癌组织中YAP阳性程度呈正相关.YAP m R N A在胰腺癌组织中的表达较正常胰腺组织明显升高(0.3682±0.0783 vs 0.0394±0.0091,P<0.01).结论:Hippo-YAP通路可能在胰腺癌的发生、发展中发挥重要作用.并有可能成为胰腺癌治疗中新的靶点.
文摘目的三阴性乳腺癌作为乳腺癌的特殊类型,尚缺乏有效的靶向治疗手段。文中旨在观察溶血磷脂酸(LPA)对三阴性乳腺癌的影响,探讨LPA-Hippo-Yes相关蛋白(YAP)信号通路在促进三阴性乳腺癌侵润转移中的作用及机制。方法体外合成YAP序列特异性小干扰RNA,应用质粒转染MDA-MB-231乳腺癌细胞株为实验组,非特异性siRNA转染MDA-MB-231细胞为阳性对照组,未转染MDA-MB-231细胞为空白对照组,每组设2个平行孔。检测各组对Hippo-YAP信号的影响,其作用机制及对Hippo-YAP上、下游信号通路调控。结果实验组总蛋白中YAP含量、细胞转染48h后的侵袭、转移能力[(0.035±0.005)、(2.200±1.000)个、(3.500±0.800)个]较阳性对照组[(0.343±0.012)、(27.600±5.100)个、(22.300±5.000)个]、空白对照组[(0.384±0.017)、(26.500±4.800)个、(22.350±6.000)个]明显降低(P<0.05)。实验组60、120、240 min时YAP表达量较阳性对照组、空白对照组显著降低(P<0.05)。实验组10、20、50μmol/L的YAP相对表达量显著低于阳性对照组、空白对照组(P<0.05)。C3转移酶和Y27632分别干预后,实验组p YAP mRNA含量(0.255±0.052、0.326±0.017)与空白对照组((0.048±0.032、0.534±0.017)、阳性对照组(0.052±0.021、0.528±0.024)比较差异有统计学意义(P<0.05)。实验组YAP m NA、AREG m NA的表达量(0.176±0.032、0.263±0.008)较空白对照组(0.043±0.013、0.051±0.014)、阴性对照组(0.049±0.025、0.057±0.043)显著升高(P<0.05)。结论 LPA诱导乳腺癌细胞侵袭转移,具有YAP蛋白依赖性,并具有时间与浓度依赖的特点;LPA-Hippo-YAP信号传导通路可能是促进三阴性乳腺癌细胞长时间转移的机制之一。
