实时监测高通量低能量激光照射诱导GSK-3β的核质穿梭  

Real-Time Monitoring of High Fluence Low-power Laser Irradiation Induced-GSK-3β Nucleocytoplasmic Shuttling

作　　者：黄磊[1] 邢达[1] 吴胜男[1] 刘镭[1] 

机构地区：[1]华南师范大学激光生命科学研究所暨激光生命科学教育部重点实验室,广东广州510631

出　　处：《激光生物学报》2009年第6期711-715,共5页Acta Laser Biology Sinica

基　　金：国家自然科学基金项目(30870676;30870658);教育部"长江学者与创新团队计划"创新团队项目(IRT0829);广东省自然科学基金项目(7117865)

摘　　要：糖原合成酶激酶-3(glycogen synthasekinase-3,GSK-3)是一种丝/苏氨酸蛋白激酶。哺乳动物细胞中主要存在GSK-3α与GSK-3β两种亚型。以前的研究认为GSK-3是一种单一的磷酸化糖原合成酶的激酶,可以抑制糖原的合成。最近的研究表明GSK-3可以磷酸化50多种底物,进而调节细胞的多种生理过程,包括细胞结构的改变、代谢,基因表达及细胞凋亡。本文主要研究在高通量低能量激光(high fluencelow-power laserirradiation,HF-LPLI)照射下,GSK-3β在活细胞中的动态分布变化。应用荧光蛋白融合蛋白GFP-GSK-3β,在人神经胶质母细胞瘤细胞(U-87)中实时监测高通量低能量激光照射下GSK-3β的动态行为。实验结果显示,120 J/cm^2的氦氖激光照射后,GSK-3β在9 h时进入细胞核,并维持在核内近2 h,随后GSK-3β又从细胞核转位到细胞质中。这表明高通量低能量激光照射激活了GSK-3β。同时,实验结果预示了高通量低能量激光照射可能激活GSK-3β并且参与调控了p53、β-catenin、Myc等相关的转录因子。进一步的研究将探讨在高通量低能量激光照射下,GSK-3β具体调控的转录因子以及调控机制。Glycogen synthase kinase-3 （ GSK-3 ） is a ubiquitous serine/threonine kinase which consists of two distinct isoforms, α and β, in mammals. Previous studies revealed that GSK-3 was only a protein kinase which could phosphorylate glycogen synthase and inhibit the glycogen kinase. Recent studies showed that GSK-3 could phosphorylate 50 kinds of substrates and regulate multiple bioprocesses, including changes of cell structure, metabolism,gene expression, and cell apoptosis. For example, heat shock could induce GSK-3β nuclear translocation to nucleus and phosphorylization of heat shock factor-1 （ HSF-1 ） , regulating apoptosis. Recent years, some studies showed that high fluence low-power la-ser irradiation （HF-LPLI） could trigger cell apoptosis, however, whether GSK-3β functioned in the process and the regulation mechanism of GSK-3β was not clear. In the present studies, we mainly investigated the changes of subcellular distribution of GSK-3β after HF-LPLI treatment in living U-87 cells using GFP-GSK-315. We found that GSK-3β translocared to nucleus at 9 hour and maintained in nucleus for nearly 2 hours, and then back to cytosol post-HF-LPLI treatment （ 120 J/cm^2）. The results indicated that GSK-3β was activated by HF-LPLI and might regulate some transcription factors, such as p53, β-catenin, and Myc. Great efforts will be made to uncover the specific kinds of transcription factors regulated by GSK-3β, and the mechanisms involved.

关 键 词：CSK-3β 细胞凋亡 高通量低能量激光照射 转录因子 

分 类 号：Q631[生物学—生物物理学]