JNK在Rap2介导的细胞转化中的功能研究

发布时间：2018-03-08 08:34

  本文选题：Rap2　切入点：JNK　出处：《哈尔滨工业大学》2010年硕士论文　论文类型：学位论文

【摘要】： Rap2属于Ras家族小分子量GTP结合蛋白家族成员之一,调节细胞的增殖、粘附、迁移、侵袭等。Rap2的功能通过其下游信号分子传递如MAP4K4,TNIK,Raf,PARG是最近发现的Rap2特异效应因子。其中MAP4K4属于STE激酶家族成员,在多种癌细胞系中促进细胞的迁移运动。而JNK作为MAP4K4,TNIK的下游效应因子,参与细胞的运动,凋亡等。在静息状态下,JNK大多以非活性的形式存在于细胞质中,一旦被激活则转移至细胞核或细胞膜调控细胞的凋亡及运动等。但JNK是否作为Rap2的下游效应因子调控细胞的迁移运动还未知。 本文通过检测Rap2对JNK表达及活性的影响及过表达Rap2对JNK在细胞内分布的影响来初步探讨Rap2-JNK信号通路在细胞迁移中的信号途径。我们发现稳定表达Rap2的HEK293细胞中,发生酪氨酸磷酸化的JNK水平较对照组明显升高。同时发现过表达Rap2-12V(Rap2的活性突变体)的细胞中JNK2的表达降低。另外,利用免疫荧光技术发现过表达Rap2的NIH3T3中JNK较对照组相比具有细胞核内聚集倾向,进一步提示Rap2对JNK的活化作用。在细胞迁移实验中,过表达Rap2的Caki-1细胞较对照组迁移速度明显加快,而加入JNK抑制剂SP600125能抑制Rap2诱导的细胞迁移。总上所述,我们的实验结果初步表明Rap2通过对JNK活性和细胞定位的调控调节细胞的迁移。
[Abstract]:Rap2 is a member of small molecular weight GTP binding protein family of Ras family, which regulates cell proliferation, adhesion and migration. The function of Rap2 is transmitted through its downstream signaling molecules, such as MAP4K4, TNIKPERG, a recently discovered Rap2 specific effector, in which MAP4K4 is a member of the STE kinase family. JNK, as a downstream effector of MAP4K4tTNIK, participates in cell movement, apoptosis and so on. In resting state, JNK exists in cytoplasm in the form of inactivity. Once activated, it was transferred to the nucleus or cell membrane to regulate the apoptosis and movement of cells, but it is not known whether JNK acts as a downstream effector of Rap2 to regulate the migration of cells. The effect of Rap2 on the expression and activity of JNK and the effect of overexpression of Rap2 on the distribution of JNK in cells were studied to explore the signal pathway of Rap2-JNK signaling pathway in cell migration. We found that the expression of Rap2 was stable in HEK293 cells. The level of JNK with tyrosine phosphorylation was significantly higher than that of control group. At the same time, the expression of JNK2 was decreased in the active mutants that expressed Rap2-12V(Rap2. It was found by immunofluorescence technique that JNK in NIH3T3, which was overexpressed by Rap2, had a tendency of aggregation in nucleus compared with that in control group, which further indicated the activation of JNK by Rap2. The migration rate of Caki-1 cells with overexpression of Rap2 was significantly faster than that of control group, and the addition of JNK inhibitor SP600125 could inhibit the cell migration induced by Rap2. Our results suggest that Rap2 regulates cell migration through regulation of JNK activity and cellular localization.
【学位授予单位】：哈尔滨工业大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329

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本文编号：1583151