细胞骨架调节蛋白3(diaphanous related formin3,DIAPH3)参与肌动蛋白重塑和调节细胞的运动和黏附,在多种肿瘤发生发展中发挥重要作用,但其在食管鳞状细胞癌(简称:食管鳞癌)中作用尚未见报道。本研究探究DIAPH3在食管鳞癌中的作用及其...细胞骨架调节蛋白3(diaphanous related formin3,DIAPH3)参与肌动蛋白重塑和调节细胞的运动和黏附,在多种肿瘤发生发展中发挥重要作用,但其在食管鳞状细胞癌(简称:食管鳞癌)中作用尚未见报道。本研究探究DIAPH3在食管鳞癌中的作用及其分子机制。首先,基因表达谱交互分析(gene expression profiling interactive analysis,GEPIA)数据库和免疫组织化学染色法,用于分析食管鳞癌组织中DIAPH3在转录水平和蛋白质水平的表达。结果显示,食管鳞癌组织中DIAPH3mRNA和蛋白质的表达高于癌旁组织,其与肿瘤分化和淋巴结转移密切相关(P<0.05)。利用Western印迹检测食管鳞癌组织中DIAPH3蛋白的表达量。结果表明:与癌旁组织相比,食管鳞癌组织中蛋白质的表达量升高(P<0.05)。再利用CCK-8、划痕愈合实验和Transwell侵袭实验,分别检测细胞的增殖、迁移和侵袭能力。实验结果显示:与对照组相比,干扰DIAPH3组细胞的增殖、迁移和侵袭能力下降(P<0.05),而过表达DIAPH3组细胞的增殖、迁移和侵袭能力增强(P<0.05)。最后,利用Western印迹检测过表达/干扰DIAPH3细胞中的细胞周期蛋白D1(cyclin D1)、E-钙黏着蛋白(E-cadherin)和波形蛋白(vimentin)的表达。Western印迹结果显示:与对照组相比,干扰DIAPH3组细胞中,E-钙黏着蛋白表达增加,且波形蛋白和细胞周期蛋白D1表达减少(P<0.05),而过表达DIAPH3组细胞中,E-钙黏着蛋白表达减少,且波形蛋白和细胞周期蛋白D1表达增加(P<0.05)。这些研究表明,DIAPH3促进食管鳞癌细胞的增殖、迁移和侵袭。其结果为食管鳞癌的诊治提供了新思路。展开更多
目的:观察应激是否损害吗啡奖赏记忆再巩固及其对海马活性调节细胞骨架蛋白(Arc)的影响。方法:(1)建立大鼠吗啡条件位置偏爱(conditioned place preference,CPP)模型,给予不同的处理即暴露、应激、暴露后立即应激,观察应激对吗啡奖赏记...目的:观察应激是否损害吗啡奖赏记忆再巩固及其对海马活性调节细胞骨架蛋白(Arc)的影响。方法:(1)建立大鼠吗啡条件位置偏爱(conditioned place preference,CPP)模型,给予不同的处理即暴露、应激、暴露后立即应激,观察应激对吗啡奖赏记忆再巩固的作用。(2)建立吗啡CPP模型,暴露6 h后应激,观察应激损害吗啡奖赏记忆再巩固是否存在时间窗。(3)处理测试后的大鼠断头取脑,采用免疫荧光法和Western blot法分析应激对吗啡CPP大鼠海马Arc表达的影响。结果:(1)吗啡奖赏记忆唤起后立即给予应激,其再巩固受到损害(P<0.05)。(2)应激损害吗啡奖赏记忆再巩固作用存在时间窗(P<0.05)。(3)暴露后给予应激组大鼠在CPP测试后海马CA1脑区中的Arc表达下降(P<0.05)。结论:(1)应激可损害吗啡奖赏记忆再巩固,且其作用具有时间窗。(2)应激损害吗啡记忆再巩固可能与其降低海马CA1脑区的Arc表达有关。展开更多
It is known that cytoskeleton-dependent trafficking of cell wall and membrane components to apical plasma membrane （PM） coupled with ion transport across pollen PM is crucial for maintaining polar pollen tube growth...It is known that cytoskeleton-dependent trafficking of cell wall and membrane components to apical plasma membrane （PM） coupled with ion transport across pollen PM is crucial for maintaining polar pollen tube growth. To elucidate whether plant hormones are involved in these processes, the effects of exogenous phytohormones, indole-3-acetic acid （IAA）, abscisic acid （ABA）, gibberellin A3 （GA3） and cytokinin （kinetin） on the growth, PM polarization, actin cytoskeleton （AC） organization and cytoplasmic pH （pile） of in vitro 4 h-growing petunia pollen tubes were investigated. IAA, ABA and GA3 displayed the growth-stimulating effects and these were accompanied by orthovanadate-sensitive hyperpolarization of the PM. Fluorescent labeling the enzyme with H＋-ATPase antibodies exhibited IAA- and ABA-induced lateral PM redistribution of it into the subapical zone of pollen tube PM. Pollen cultivation on the medium with latrunculin B, the inhibitor of actin polymerization, resulted in inhibition of pollen tube growth and simultaneously in the drop of endogenous IAA content. The IAA-growth stimulating effect was correlated with increased content of actin filaments （AF） in both apical and subapical zones of tubes, while ABA and GA3 exerted the same effect but it was accompanied by redistributing F-actin only to apical zone. In contrast, kinetin decreased the total F-actin content and inhibited pollen tube growth. It has been shown that the pHe of growing pollen tubes is sensitive to the plant hormones. In the case of male gametophyte growing for 1, 2 and 4 h, IAA induced alkalinization of the cytosol, while ABA and GA3 exerted qualitatively similar effect only after its growth for 1 h and 4 h, respectively. Kinetin, in contrast, resulted in acidification of the cytosol. All these results, taken together, indicate, for the first time, potential targets of the phytohormone action in pollen tubes.展开更多
文摘目的:观察应激是否损害吗啡奖赏记忆再巩固及其对海马活性调节细胞骨架蛋白(Arc)的影响。方法:(1)建立大鼠吗啡条件位置偏爱(conditioned place preference,CPP)模型,给予不同的处理即暴露、应激、暴露后立即应激,观察应激对吗啡奖赏记忆再巩固的作用。(2)建立吗啡CPP模型,暴露6 h后应激,观察应激损害吗啡奖赏记忆再巩固是否存在时间窗。(3)处理测试后的大鼠断头取脑,采用免疫荧光法和Western blot法分析应激对吗啡CPP大鼠海马Arc表达的影响。结果:(1)吗啡奖赏记忆唤起后立即给予应激,其再巩固受到损害(P<0.05)。(2)应激损害吗啡奖赏记忆再巩固作用存在时间窗(P<0.05)。(3)暴露后给予应激组大鼠在CPP测试后海马CA1脑区中的Arc表达下降(P<0.05)。结论:(1)应激可损害吗啡奖赏记忆再巩固,且其作用具有时间窗。(2)应激损害吗啡记忆再巩固可能与其降低海马CA1脑区的Arc表达有关。
文摘It is known that cytoskeleton-dependent trafficking of cell wall and membrane components to apical plasma membrane （PM） coupled with ion transport across pollen PM is crucial for maintaining polar pollen tube growth. To elucidate whether plant hormones are involved in these processes, the effects of exogenous phytohormones, indole-3-acetic acid （IAA）, abscisic acid （ABA）, gibberellin A3 （GA3） and cytokinin （kinetin） on the growth, PM polarization, actin cytoskeleton （AC） organization and cytoplasmic pH （pile） of in vitro 4 h-growing petunia pollen tubes were investigated. IAA, ABA and GA3 displayed the growth-stimulating effects and these were accompanied by orthovanadate-sensitive hyperpolarization of the PM. Fluorescent labeling the enzyme with H＋-ATPase antibodies exhibited IAA- and ABA-induced lateral PM redistribution of it into the subapical zone of pollen tube PM. Pollen cultivation on the medium with latrunculin B, the inhibitor of actin polymerization, resulted in inhibition of pollen tube growth and simultaneously in the drop of endogenous IAA content. The IAA-growth stimulating effect was correlated with increased content of actin filaments （AF） in both apical and subapical zones of tubes, while ABA and GA3 exerted the same effect but it was accompanied by redistributing F-actin only to apical zone. In contrast, kinetin decreased the total F-actin content and inhibited pollen tube growth. It has been shown that the pHe of growing pollen tubes is sensitive to the plant hormones. In the case of male gametophyte growing for 1, 2 and 4 h, IAA induced alkalinization of the cytosol, while ABA and GA3 exerted qualitatively similar effect only after its growth for 1 h and 4 h, respectively. Kinetin, in contrast, resulted in acidification of the cytosol. All these results, taken together, indicate, for the first time, potential targets of the phytohormone action in pollen tubes.
