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模拟微重力诱导的细胞微丝变化影响{sl 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。该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#alpha#1链基因(collagen type I alpha chain 1 gene，{sl 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{sl 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{sl 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{sl 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;

Disruption of microfilament cytoskeleton induced by simulated microgravity increases the activity of {sl COL1A1} promoter

Dai Zhongquan, Li Yinghui, Ding Bai, Yang Fen, Tan Yingjun, Nie Jielin, Yu Jianru

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 Photon-M1 or clinorotation simulated microgravity, which suggested that microgravity destroyed the well-order cell cytoskeleton and induced a rearrangement. Treatment with suitable concentration of cytochalasin B in normal gravity induced disruption of microfilament, increased the activity of COL1A1 promoter and resulted in a dose-dependent increase of EGFP fluorescence. Therefore, a certain extent disruption of the microfilament system was associated with increased activity of the COL1A1 promoter. All above demonstrate that microfilament cytoskeleton system takes part in the regulation of COL1A1 promoter activity and plays an important role in the signaling of microgravity.

Key words: Microgravity;Cytoskeleton;Promoter;Collagen type I ;

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引用本文：

戴钟铨, 李莹辉, 丁柏, 杨芬, 谭映军, 聂捷琳, 于建如 . 模拟微重力诱导的细胞微丝变化影响{sl COL1A1}启动子活性[J]. 生理学报 2006; 58 (1): .

Dai Zhongquan, Li Yinghui, Ding Bai, Yang Fen, Tan Yingjun, Nie Jielin, Yu Jianru. Disruption of microfilament cytoskeleton induced by simulated microgravity increases the activity of {sl COL1A1} promoter. Acta Physiol Sin 2006; 58 (1): (in Chinese with English abstract).