Objective: To study the chemotactic superfamily genes expression profiling of macrophage line U937 treated with monocyte chemoattractant protein-1 (MCP-1) using gene chip technique. Methods: Total RNA from macrophage ...Objective: To study the chemotactic superfamily genes expression profiling of macrophage line U937 treated with monocyte chemoattractant protein-1 (MCP-1) using gene chip technique. Methods: Total RNA from macrophage line U937 (as control) and U937 with MCP-1 was extracted, made reverse transcript to cDNA and tested with gene expression chip HO2 human. Results: Some chemotactic-related gene expressions were changed in all analyzed genes. Regulated upon activation, normal T cell expressed and secreted (RANTES) was up-regulated over 2-fold and 7 chemotactic-related genes (CCR2, CCR5, CCL16, GROβ, GROγ, IL-8 and granulocyte chemotactic protein 2) were down-regulated over 2-fold in MCP-1 treated U937 cells at mRNA level. Conclusion: MCP-1 can influence some chemokines and receptors expression in macrophage in vitro, in which MCP-1 mainly down-regulates the chemotactic genes expression of those influencing neutrophilic granulocyte (GROβ, GROγ, IL-8 and granulocyte chemotactic protein 2). Another novel finding is that it can also down-regulate the mRNA level of CCR5, which plays a critical role in many disorders and illnesses.展开更多
目的:对HL60细胞在NSC67657作用下向单核系分化前后,双向电泳分离的表达差异蛋白β-catenin相关蛋白1(Beta-catenin-interacting protein 1,ICAT)进行验证,并对ICAT在细胞分化中的功能进行研究.方法:通过RT-PCR和Western blot方法验证...目的:对HL60细胞在NSC67657作用下向单核系分化前后,双向电泳分离的表达差异蛋白β-catenin相关蛋白1(Beta-catenin-interacting protein 1,ICAT)进行验证,并对ICAT在细胞分化中的功能进行研究.方法:通过RT-PCR和Western blot方法验证药物作用细胞前后ICAT基因和蛋白的表达差异;通过免疫荧光协同分析目的蛋白的表达水平,并对其进行初步定位.构建pDsRed-ICAT真核表达载体,转染HL60细胞,筛选阳性克隆.对ICAT基因重组质粒转染细胞作细胞形态学、细胞增殖改变的观察和细胞周期检测以及超微结构观察.结果:NSC67657诱导HL60细胞向单核系分化,ICAT蛋白表达上调,其主要定位于细胞核和胞质.真核表达载体构建成功,电转后G418筛选可得90%以上阳性克隆.转染重组质粒的HL60细胞增殖受抑,电镜下胞核异染色质密集,核质比减小,表面抗原CD14表达和对照组无差异,但在药物处理后24h即可表达71.3%,明显高于对照组,瑞氏染色可见明显分化细胞.结论:ICAT蛋白在NSC67657诱导HL60细胞分化中表达上调,但仅是过表达的ICAT基因并不能诱导HL60细胞向单核系分化,却能提高HL60细胞对NSC67657诱导作用的敏感性.展开更多
文摘Objective: To study the chemotactic superfamily genes expression profiling of macrophage line U937 treated with monocyte chemoattractant protein-1 (MCP-1) using gene chip technique. Methods: Total RNA from macrophage line U937 (as control) and U937 with MCP-1 was extracted, made reverse transcript to cDNA and tested with gene expression chip HO2 human. Results: Some chemotactic-related gene expressions were changed in all analyzed genes. Regulated upon activation, normal T cell expressed and secreted (RANTES) was up-regulated over 2-fold and 7 chemotactic-related genes (CCR2, CCR5, CCL16, GROβ, GROγ, IL-8 and granulocyte chemotactic protein 2) were down-regulated over 2-fold in MCP-1 treated U937 cells at mRNA level. Conclusion: MCP-1 can influence some chemokines and receptors expression in macrophage in vitro, in which MCP-1 mainly down-regulates the chemotactic genes expression of those influencing neutrophilic granulocyte (GROβ, GROγ, IL-8 and granulocyte chemotactic protein 2). Another novel finding is that it can also down-regulate the mRNA level of CCR5, which plays a critical role in many disorders and illnesses.
文摘目的:对HL60细胞在NSC67657作用下向单核系分化前后,双向电泳分离的表达差异蛋白β-catenin相关蛋白1(Beta-catenin-interacting protein 1,ICAT)进行验证,并对ICAT在细胞分化中的功能进行研究.方法:通过RT-PCR和Western blot方法验证药物作用细胞前后ICAT基因和蛋白的表达差异;通过免疫荧光协同分析目的蛋白的表达水平,并对其进行初步定位.构建pDsRed-ICAT真核表达载体,转染HL60细胞,筛选阳性克隆.对ICAT基因重组质粒转染细胞作细胞形态学、细胞增殖改变的观察和细胞周期检测以及超微结构观察.结果:NSC67657诱导HL60细胞向单核系分化,ICAT蛋白表达上调,其主要定位于细胞核和胞质.真核表达载体构建成功,电转后G418筛选可得90%以上阳性克隆.转染重组质粒的HL60细胞增殖受抑,电镜下胞核异染色质密集,核质比减小,表面抗原CD14表达和对照组无差异,但在药物处理后24h即可表达71.3%,明显高于对照组,瑞氏染色可见明显分化细胞.结论:ICAT蛋白在NSC67657诱导HL60细胞分化中表达上调,但仅是过表达的ICAT基因并不能诱导HL60细胞向单核系分化,却能提高HL60细胞对NSC67657诱导作用的敏感性.
