机械变形培养皿对大鼠骨肉瘤细胞cAMP和DNA合成的影响。

Y J Chen, L T Hou, H F Chang, K C Chen
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摘要

本研究的目的是记录大鼠骨肉瘤细胞(ROS 17/2.8)对体外机械应力的生化反应。采用应力施加装置研究机械应力对ROS 17/2.8细胞的影响。简单地说,该装置是通过在塑料培养皿底部外用自固化丙烯酸树脂粘接正畸膨胀螺钉制成的。通过激活膨胀螺钉使培养皿产生不可逆的变形。由此产生的机械应力被转移到附着在培养皿上的细胞上。采用竞争性蛋白结合法测定了ROS 17/2.8细胞对机械应力的环磷酸腺苷(cAMP)反应。机械应力对细胞生长的影响通过加入3h -胸腺嘧啶在不同时期的机械应力施加后进行评估。结果表明,机械应力可使应激刺激后早期ROS 17/2.8细胞cAMP的生成增加。这种cAMP水平的变化取决于施加应力的持续时间,当施加机械应力一小时时发生最大响应。虽然在机械应力作用1小时的ROS 17/2.8细胞中,3h -胸苷的细胞掺入减少了40-60%,但当机械应力持续24小时时,该反应从抑制作用恢复到基线的80-85%。本研究的观察结果表明,机械应力刺激cAMP的产生,并抑制应激早期ROS 17/2.8细胞的3h -胸腺嘧啶掺入。
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The response of cAMP and DNA synthesis in rat osteosarcoma cells to mechanically deformed culture dishes.

The purpose of this research was to document the biochemical response of rat osteosarcoma cells (ROS 17/2.8) to mechanical stress in vitro. The influence of mechanical stress on ROS 17/2.8 cells was studied using a stress application device. Briefly, the device was fabricated by bonding an orthodontic expansion screw outside the bottom of a plastic culture dish with self-curing acrylic resin. Irreversible deformation of the culture dish was produced by activating an expansion screw. The resulting mechanical stress was transferred to the cells which attached to the culture dish. The response in terms of cyclic adenosine monophosphate (cAMP) of ROS 17/2.8 cells to mechanical stress was measured using a competitive protein-binding method. The effect of mechanical stress on cellular growth was assessed through the incorporation of 3H-thymidine after different periods of mechanical stress application. The results revealed that mechanical stress could increase the production of cAMP in ROS 17/2.8 cells at an early phase after stress stimulation. This change in the cAMP level was dependent on the duration of stress application, and the maximal response occurred when the mechanical stress was applied for one hour. Although the cellular incorporation of 3H-thymidine decreased 40-60% in ROS 17/2.8 cells subjected to mechanical stress for 1 hour, this reaction recovered from the inhibition effect to 80-85% of the baseline when the mechanical stress lasted for 24 hours. The observations in this study indicate that mechanical stress stimulates the production of cAMP and inhibits the 3H-thymidine incorporation of ROS 17/2.8 cells at an early phase of stress application.

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