目的探讨α吡喃酮类化合物Rasfonin对鸟苷酸交换因子SOS1表达的调控作用及其机制。方法(1)将MCF-7、Calu-1和UM-UC-3细胞分别分为溶剂对照组和Rasfonin组(1、5、10和15μmol·L^(-1)),处理24 h后CCK-8法检测MCF-7、Calu-1和UM-UC-3...目的探讨α吡喃酮类化合物Rasfonin对鸟苷酸交换因子SOS1表达的调控作用及其机制。方法(1)将MCF-7、Calu-1和UM-UC-3细胞分别分为溶剂对照组和Rasfonin组(1、5、10和15μmol·L^(-1)),处理24 h后CCK-8法检测MCF-7、Calu-1和UM-UC-3细胞存活率。将MCF-7、Calu-1和UM-UC-3细胞分别分为细胞对照组(培养基)、表皮生长因子组(EGF 50μg·L^(-1)处理5 min),EGF+Rasfonin组(Rasfonin分别以5和10μmol·L^(-1)预处理不同时间后,再EGF 50μg·L^(-1)处理5 min),实时荧光定量PCR和Western印迹法检测MCF-7、Calu-1和UM-UC-3细胞中SOS1 m RNA和蛋白表达水平。(2)将阴性对照(NC)质粒、KRASWT质粒和KRASG12C质粒分别与SOS1质粒共转染至293T细胞,转染后分为溶剂对照组和Rasfonin组(1、5和10μmol·L^(-1)),处理12 h后采用双荧光素酶报告基因实验检测SOS1启动子活性。将质粒KRAS^(WT)、KRAS^(G12C)、NC+SOS1、KRAS^(WT)+SOS1和KRAS^(G12C)+SOS1分别转染至293T细胞,分为EGF组和EGF+Rasfonin组(Rasfonin 10μmol·L^(-1)预处理12 h后,再EGF处理5 min),Western印迹法检测293T细胞中SOS1蛋白表达水平。结果(1)与溶剂对照组相比,Rasfonin 5、10和15μmol·L^(-1)显著抑制Calu-1和UM-UC-3细胞增殖,IC_(50)分别为8.22和4.94μmol·L^(-1),MCF-7细胞的IC50为45.15μmol·L^(-1)。Rasfonin处理3 h MCF-7细胞SOS1 m RNA水平升高;Rasfonin处理1 h Calu-1细胞SOS1 m RNA水平升高,3和6 h SOS1 m RNA水平降低;Rasfonin处理3 h UM-UC-3细胞SOS1 m RNA表达,6 h降低。与EGF组相比,EGF+Rasfonin组MCF-7细胞的SOS1蛋白表达水平无显著变化,Calu-1与UM-UC-3细胞的SOS1蛋白表达水平显著降低。(2)与溶剂对照组相比,Rasfonin对SOS1单表达组SOS1启动子活性无显著影响,但SOS1与KRAS^(WT)或KRAS^(G12C)蛋白共表达时,SOS1启动子活性被Rasfonin显著抑制。与EGF组相比,Rasfonin对SOS1单表达组及SOS1+KRAS^(WT)共表达组的SOS1蛋白表达水平无显著变化,SOS1+KRAS^(G12C)共表达组的SOS1蛋白表达水平显著降低。结论Rasfonin通过KRASG12C依赖性途径抑制SOS1表达,这可能是其抗肿瘤作用的机制之一。展开更多
SOS1(son of sevenless homolog 1)蛋白是一种在细胞中广泛表达的调控蛋白。作为信号通路中的关键蛋白,SOS1在细胞内许多信号转导通路中起着重要的调控作用,例如参与调控Ras和Rac信号通路。SOS1的异常表达或突变与临床疾病的发生密切相...SOS1(son of sevenless homolog 1)蛋白是一种在细胞中广泛表达的调控蛋白。作为信号通路中的关键蛋白,SOS1在细胞内许多信号转导通路中起着重要的调控作用,例如参与调控Ras和Rac信号通路。SOS1的异常表达或突变与临床疾病的发生密切相关。本文对SOS1的功能及其在生理学、病理生理学中的作用的研究进展进行综述。展开更多
Sweet sorghum mutants induced by^(12)C(6+)-ion irradiation were planted under different soil salinity conditions to investigate the mechanisms maintaining the transport and spatial distribution of Na^+. The functions ...Sweet sorghum mutants induced by^(12)C(6+)-ion irradiation were planted under different soil salinity conditions to investigate the mechanisms maintaining the transport and spatial distribution of Na^+. The functions of the synergistic responses of NHX, AKT1, and SOS1 related to Na^+ accumulation were investigated in control(KFJT-CK) sorghum and KF1210-3 and KF1210-4 mutants. The results indicated that the NHX, AKT1, and SOS1 proteins in sweet sorghum are mainly involved in the transport, exclusion, and spatial distribution of Na^+,respectively. In addition to physiological parameters, we also measured the expression levels of NHX, AKT1, and SOS1 genes. The experimental results indicated that 150 m M Na Cl induced marked increases in the transcripts of NHX and SOS1 after 8 and 12 h in the KF1210-3,KF1210-4, and KFJT-CK cultivars. In contrast, however, a decrease in AKT1 was observed. On the basis of our results, we propose a model in which cooperation amongNHX, AKT1, and SOS1 facilitates Na^+ homeostasis in sweet sorghum in response to an increase in salt concentration. Accordingly, study of the regulatory mechanisms in sweet sorghum generated by carbon ion irradiation is essential for the selection of salt-tolerant cultivars.展开更多
文摘目的探讨α吡喃酮类化合物Rasfonin对鸟苷酸交换因子SOS1表达的调控作用及其机制。方法(1)将MCF-7、Calu-1和UM-UC-3细胞分别分为溶剂对照组和Rasfonin组(1、5、10和15μmol·L^(-1)),处理24 h后CCK-8法检测MCF-7、Calu-1和UM-UC-3细胞存活率。将MCF-7、Calu-1和UM-UC-3细胞分别分为细胞对照组(培养基)、表皮生长因子组(EGF 50μg·L^(-1)处理5 min),EGF+Rasfonin组(Rasfonin分别以5和10μmol·L^(-1)预处理不同时间后,再EGF 50μg·L^(-1)处理5 min),实时荧光定量PCR和Western印迹法检测MCF-7、Calu-1和UM-UC-3细胞中SOS1 m RNA和蛋白表达水平。(2)将阴性对照(NC)质粒、KRASWT质粒和KRASG12C质粒分别与SOS1质粒共转染至293T细胞,转染后分为溶剂对照组和Rasfonin组(1、5和10μmol·L^(-1)),处理12 h后采用双荧光素酶报告基因实验检测SOS1启动子活性。将质粒KRAS^(WT)、KRAS^(G12C)、NC+SOS1、KRAS^(WT)+SOS1和KRAS^(G12C)+SOS1分别转染至293T细胞,分为EGF组和EGF+Rasfonin组(Rasfonin 10μmol·L^(-1)预处理12 h后,再EGF处理5 min),Western印迹法检测293T细胞中SOS1蛋白表达水平。结果(1)与溶剂对照组相比,Rasfonin 5、10和15μmol·L^(-1)显著抑制Calu-1和UM-UC-3细胞增殖,IC_(50)分别为8.22和4.94μmol·L^(-1),MCF-7细胞的IC50为45.15μmol·L^(-1)。Rasfonin处理3 h MCF-7细胞SOS1 m RNA水平升高;Rasfonin处理1 h Calu-1细胞SOS1 m RNA水平升高,3和6 h SOS1 m RNA水平降低;Rasfonin处理3 h UM-UC-3细胞SOS1 m RNA表达,6 h降低。与EGF组相比,EGF+Rasfonin组MCF-7细胞的SOS1蛋白表达水平无显著变化,Calu-1与UM-UC-3细胞的SOS1蛋白表达水平显著降低。(2)与溶剂对照组相比,Rasfonin对SOS1单表达组SOS1启动子活性无显著影响,但SOS1与KRAS^(WT)或KRAS^(G12C)蛋白共表达时,SOS1启动子活性被Rasfonin显著抑制。与EGF组相比,Rasfonin对SOS1单表达组及SOS1+KRAS^(WT)共表达组的SOS1蛋白表达水平无显著变化,SOS1+KRAS^(G12C)共表达组的SOS1蛋白表达水平显著降低。结论Rasfonin通过KRASG12C依赖性途径抑制SOS1表达,这可能是其抗肿瘤作用的机制之一。
基金These authors contributed equally to this review.
文摘SOS1(son of sevenless homolog 1)蛋白是一种在细胞中广泛表达的调控蛋白。作为信号通路中的关键蛋白,SOS1在细胞内许多信号转导通路中起着重要的调控作用,例如参与调控Ras和Rac信号通路。SOS1的异常表达或突变与临床疾病的发生密切相关。本文对SOS1的功能及其在生理学、病理生理学中的作用的研究进展进行综述。
基金supported by the Science and Technology Service Network Initiative(STS)program of the Chinese Academy of Sciences(CAS)(KFJ-EW-STS-086)the National Natural Science Foundation of China(No.11275171)+1 种基金the CAS‘‘Light of West China’’ Program(Nos.29Y506020 and 29Y406020)the Youth Innovation Promotion Association of CAS(No.2015337)
文摘Sweet sorghum mutants induced by^(12)C(6+)-ion irradiation were planted under different soil salinity conditions to investigate the mechanisms maintaining the transport and spatial distribution of Na^+. The functions of the synergistic responses of NHX, AKT1, and SOS1 related to Na^+ accumulation were investigated in control(KFJT-CK) sorghum and KF1210-3 and KF1210-4 mutants. The results indicated that the NHX, AKT1, and SOS1 proteins in sweet sorghum are mainly involved in the transport, exclusion, and spatial distribution of Na^+,respectively. In addition to physiological parameters, we also measured the expression levels of NHX, AKT1, and SOS1 genes. The experimental results indicated that 150 m M Na Cl induced marked increases in the transcripts of NHX and SOS1 after 8 and 12 h in the KF1210-3,KF1210-4, and KFJT-CK cultivars. In contrast, however, a decrease in AKT1 was observed. On the basis of our results, we propose a model in which cooperation amongNHX, AKT1, and SOS1 facilitates Na^+ homeostasis in sweet sorghum in response to an increase in salt concentration. Accordingly, study of the regulatory mechanisms in sweet sorghum generated by carbon ion irradiation is essential for the selection of salt-tolerant cultivars.
