模拟微重力诱导细胞微丝变化影响COL1A1启动子活性.pdfVIP

生理学报 Acta Physiologica Sinica, February 25, 2006, 58 (1): 53-57 53 研究论文 模拟微重力诱导的细胞微丝变化影响COL1A1 启动子活性 * 戴钟铨，李莹辉 ，丁柏，杨芬，谭映军，聂捷琳，于建如 航天细胞分子生物学实验室，航天医学工程研究所，北京100094 摘 要：细胞骨架系统是细胞内的重力感受系统。已知微重力导致的细胞形态、功能、信号传导等多种变化均与细胞骨 架系统变化有关，但微重力对相关基因调控的影响知之甚少。本研究以构建的基因工程细胞株(EGFP-ROS) 为对象，以回 转器模拟微重力效应，利用增强型绿色荧光蛋白(enhanced green fluorescence protein ，EGFP)荧光半定量和细胞微丝荧光染 色分析技术，探讨回转模拟微重力条件下，细胞微丝系统对I 型胶原α1 链基因(collagen type I alpha chain 1 gene，COL1A1) 启动子活性的影响。空间飞行和回转模拟微重力后，细胞微丝解聚、张力纤维减少，表明微重力可降低细胞微丝结构的 有序性，诱导细胞骨架重排。适合剂量的细胞松弛素B 处理EGFP-ROS 细胞诱导微丝骨架解聚，同时导致COL1A1 启动 子活性增加，细胞荧光强度增强，并呈现剂量依赖性。因此，一定程度的细胞微丝系统破坏将导致COL1A1 启动子活性 的增强，证明细胞微丝骨架系统参与了微重力对COL1A1 启动子活性调节，且在微重力信号传导中起重要作用。 关键词：微重力；细胞骨架；启动子；I 型胶原 中图分类号：R852.22 Disruption of microfilament cytoskeleton induced by simulated microgravity increases the activity of COL1A1 promoter * DAI Zhong-Quan, LI Ying-Hui , DING Bai, YANG Fen, TAN Ying-Jun, NIE Jie-Lin, YU Jian-Ru Laboratory of Space Cellular and Molecular Biology, Institute of Space Medico-engineering, Beijing 100094, China Abstract: It is well known that cytoskeleton system is the sensor of gravity in cells. Under microgravity condition, cytoskeleton is associated with the changes of cell shape, function, signaling and so on; but the relationship between cytoskeleton and gene expression is not fully understood. In present study, we discussed the effects of cell microfilament on the activity of collagen type I alpha 1 chain gene (COL1A1) promoter under microgravity simulated by clinostat and/or cytochalasin B as microfilament depolymerizer in the established EGFP-ROS cell line using the method of fluorescence semi-quantitative analysis and the fluorescent stain of microfilament. Compared with the normal control, the microfilament of ROS17/2.8 cell tended to disassemble, marginal distribution of fiber stress, and showed reducing stress fibers after spaceflight in P