Cyclic stretch-induced the cytoskeleton rearrangement and gene expression of cytoskeletal regulators in human periodontal ligament cells
Cyclic stretch-induced the cytoskeleton rearrangement and gene expression of cytoskeletal regulators in human periodontal ligament cells
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DOI：
10.1080/00016357.2017.1347823
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发表时间：
2017-07
影响因子：
2
通讯作者：
Yaqin Wu;Jiabao Zhuang;Dan Zhao;Fuqiang Zhang;Jiayin Ma;Chun Xu
中科院分区：
文献类型：
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作者：
Yaqin Wu;Jiabao Zhuang;Dan Zhao;Fuqiang Zhang;Jiayin Ma;Chun Xu
摘要目的：本研究旨在通过分离人PDL细胞中与细胞骨架调节相关的几个力学反应基因，探讨牵张诱导细胞重排和细胞骨架重排的机制。材料和方法：用1、10、20%的应变分别拉伸细胞0.5、1、2、4、6、12、24 h后，记录并计算微丝形态和含量的变化。同时，用实时荧光定量聚合酶链式反应芯片检测细胞骨架调节因子编码基因在20%菌株作用6 h和24 h后的表达。Western印迹分析这些差异表达基因编码的几种细胞骨架调节因子的蛋白表达水平。结果：共聚焦荧光染色结果证实牵张诱导细胞重排和微丝重排。在筛选的84个基因中，1个基因在拉伸6h后表达上调，2个基因在拉伸6h后表达下调。同时，拉伸24小时后，3个基因表达上调，2个基因表达下调。这些差异表达的基因包括调控微丝聚合/解聚的基因(CDC42EP2、FNBP1L、NCK2、PIKfyve、Was1)、微管聚合/解聚基因(STMN1)、微丝与微管相互作用基因(MACF1)以及磷酸酶基因(PPP1R12B)。在这些基因编码的蛋白质中，CDC42EP2编码的CDC42效应蛋白-2和STMN1编码的stathmin-1的蛋白表达水平下调，而WAS1编码的N-WASP的蛋白表达水平上调。结论：本研究证实了周期性拉伸诱导的人PDL细胞微丝的重排和重排，并发现了一些与细胞骨架调节有关的力敏感基因。
Abstract Objective: This study aimed to explore the mechanism of the stretch-induced cell realignment and cytoskeletal rearrangement by identifying several mechanoresponsive genes related to cytoskeletal regulators in human PDL cells. Material and methods: After the cells were stretched by 1, 10 and 20% strains for 0.5, 1, 2, 4, 6, 12 or 24 h, the changes of the morphology and content of microfilaments were recorded and calculated. Meanwhile, the expression of 84 key genes encoding cytoskeletal regulators after 6 and 24 h stretches with 20% strain was detected by using real-time PCR array. Western blot was applied to identify the protein expression level of several cytoskeletal regulators encoded by these differentially expressed genes. Results: The confocal fluorescent staining results confirmed that stretch-induced realignment of cells and rearrangement of microfilaments. Among the 84 genes screened, one gene was up-regulated while two genes were down-regulated after 6 h stretch. Meanwhile, three genes were up-regulated while two genes were down-regulated after 24 h stretch. These genes displaying differential expression included genes regulating polymerization/depolymerization of microfilaments (CDC42EP2, FNBP1L, NCK2, PIKFYVE, WASL), polymerization/depolymerization of microtubules (STMN1), interacting between microfilaments and microtubules (MACF1), as well as a phosphatase (PPP1R12B). Among the proteins encoded by these genes, the protein expression level of Cdc42 effector protein-2 (encoded by CDC42EP2) and Stathmin-1 (encoded by STMN1) was down-regulated, while the protein expression level of N-WASP (encoded by WASL) was up-regulated. Conclusion: The present study confirmed the cyclic stretch-induced cellular realignment and rearrangement of microfilaments in the human PDL cells and indicated several force-sensitive genes with regard to cytoskeletal regulators.