DADS拮抗SRC通路对DJ-1核内高表达HL-60细胞侵袭迁移的影响

发布时间：2018-01-09 14:22

本文关键词：DADS拮抗SRC通路对DJ-1核内高表达HL-60细胞侵袭迁移的影响　出处：《南华大学》2015年硕士论文　论文类型：学位论文

【摘要】：[目的]研究二烯丙基二硫(DADS)对DJ-1核内高表达人白血病HL-60细胞侵袭迁移影响的作用机制,为白血病治疗的发展提供更完善的分子机制,为白血病患者找出更多的有效药物。[方法]通过Western blot检测DADS对DJ-1核内高表达HL-60细胞DJ-1表达的影响,以及DADS对DJ-1核内高表达HL-60细胞Src通路的影响。随后,再通过Western blot检测DADS及Src抑制剂对DJ-1核内高表达HL-60细胞Src通路的影响,以及DADS与Src抑制剂联合作用下对DJ-1表达的影响。应用Transwell迁移侵袭小室实验分别观察DADS及Src抑制剂对DJ-1核内高表达HL-60细胞增殖、侵袭迁移的影响。针对实验所需,实验分三组:对照组(HL-60细胞)、空载体组及高表达组(DJ-1核内高表达HL-60细胞)。[结果]Western blot结果显示,DADS处理后的DJ-1核内高表达HL-60细胞24h、48h、72h后,DJ-1蛋白表达水平表现为时间依赖性的下降(P0.05)。经DADS处理后,对照组、空载体组、高表达细胞p-Src、p-Fak的蛋白表达水平较未处理的各组均显著下降(P0.05),三组细胞的Src、Fak、Integrin的总蛋白表达水平没有显著变化。随后的Western blot结果显示,DADS及Src抑制剂处理后,对照组、空载体组、高表达组细胞的p-Src、p-Fak的蛋白表达水平较未处理的各组均显著下降(P0.05),而三组细胞Src、Fak、Integrin的总蛋白表达水平亦没有显著变化。二者联合处理DJ-1核内高表达HL-60细胞24h、48h、72h后,DJ-1蛋白水平表现为时间依赖性的下降(P0.05)。Transwell迁移和侵袭实验结果显示,DADS、Src抑制剂单独以及联合处理对照组与高表达组细胞后,DADS与抑制剂均可以抑制细胞侵袭迁移能力(P0.05),且经二者联合处理后的细胞侵袭迁移抑制作用更显著(P0.05)。[结论]:1.DADS可通过拮抗Src信号通路,下调DJ-1,抑制白血病细胞侵袭与迁移。2.Src抑制剂也具有抑制白血病细胞侵袭迁移的作用,且Src抑制剂可以增强DADS抗肿瘤的效应。
[Abstract]:[Objective] to study the diallyl two sulfur (DADS) on nuclear DJ-1 high expression mechanism in human leukemia HL-60 cells invasion and migration effect, provide a molecular mechanism for the development of better treatment of leukemia, to find effective drugs. More for leukemia patients by Western blot DADS on the detection of nuclear DJ-1 in high expression expression DJ-1 HL-60 cells, and DADS on the nuclear DJ-1 effect of high expression of HL-60 cells of the Src pathway. Then, through the Western blot detection of DADS and Src inhibitors on the nuclear DJ-1 effect of high expression of HL-60 cells in the Src pathway, and the effect on the expression of DJ-1 DADS and Src under the combined effect of preparations. The application of Transwell cell migration and invasion experimental observation of DADS and Src inhibitors on nuclear DJ-1 expression of HL-60 cell proliferation, invasion and migration. According to the experimental effect required, were divided into three groups: control (HL-60 cells), vector group and high The expression of group (DJ-1 HL-60 high expression cell nucleus). The results showed that]Western blot results, after treatment with DADS DJ-1 in the nucleus of HL-60 cells highly expressed 24h, 48h, 72h, declined for the time dependence of the performance level of the expression of DJ-1 protein (P0.05). After DADS treatment, control group, vector group, high the expression of p-Src in each group, the expression level of p-Fak protein were significantly decreased compared with untreated cells (P0.05), three groups of Src, Fak, total protein expression level of Integrin did not change significantly. Then the Western blot showed that DADS and Src inhibitor treatment, control group, empty vector group and high expression group p-Src cells, the expression level of p-Fak protein was higher than the untreated were significantly decreased (P0.05), and the three group of cells Src, Fak, total protein expression level of Integrin has no significant change. The two combined treatment of nuclear DJ-1 in high expression of HL-60 cell 24h, 48h, 72h, DJ-1 protein expression level Drop for time dependent (P0.05).Transwell migration and invasion assay showed that DADS inhibitor Src alone and combined with control group and high expression group cells, and DADS inhibitors can inhibit cell migration and invasion (P0.05), and the two combined treatment after the cell invasion and migration inhibition plays a more significant role. (P0.05). Conclusion: 1.DADS] by antagonizing Src signaling pathway and down-regulation of DJ-1, leukemia cell invasion and migration of.2.Src inhibitors also inhibit the migration and invasion of leukemia cells, and Src inhibitors can enhance the antitumor effect of DADS.

【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R733.7

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