Studying the effects of actin cytoskeletal destabilization on cell cycle by cofilin overexpression
Studying the effects of actin cytoskeletal destabilization on cell cycle by cofilin overexpression
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DOI：
10.1385/mb:31:1:001
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发表时间：
2005-09-01
影响因子：
2.6
通讯作者：
Keng, PC
中科院分区：
文献类型：
--
作者：
Lee, YJ;Keng, PC
肌动蛋白细胞骨架对细胞生长的重要性历史上是使用细胞松弛素等有毒药物来研究的。然而，这些药物的不可预测的影响可能会影响报告的观察结果。在我们的研究中，我们建立了一个利用丝切蛋白过度表达的无药物系统来研究肌动蛋白丝与细胞周期进程之间的关系。 Cofilin是肌动蛋白解聚因子（ADF）/cofilin家族的成员，将cofilin cDNA克隆至四环素诱导基因表达载体并稳定转染人肺癌H1299上皮细胞。通过肌动蛋白分析和显微镜检查，分别检测到肌丝蛋白丝切蛋白过表达细胞中肌动蛋白丝的不稳定和形态变化。生长速率的测量表明，在丝切蛋白过度表达的细胞中，细胞增殖受到抑制。此外，细胞周期分析表明，约 90% 的 cofilin 过表达细胞在 G1 期停滞，这与之前的报道一致，即药物介导的肌动蛋白丝破坏可导致 G I 期停滞。综上所述，肌动蛋白丝切蛋白过表达细胞模型提供了证据，表明可以使用分子方法而不是药物来研究肌动蛋白细胞骨架不稳定对细胞周期进程的影响。
The significance of actin cytoskeleton on cell growth was historically studied using toxic drugs, such as cytochalasin. However, it is possible that unpredictable effects of these agents may have influenced the reported observations. In our study, we have established a drug-free system using cofilin overexpression to investigate the relationship between actin filaments and cell cycle progression. Cofilin is a member of the actin depolymerization factor (ADF)/cofilin family, cofilin cDNA was cloned to a tetracycline-inducible gene expression vector and stably transfected to human lung cancer H1299 epithelial cells. Destabilization of actin filaments and morphological change was detected in cofilin overexpressing cells by actin analysis and microscopy, respectively. Measurements of growth rates showed that cell proliferation was retarded in cells with overexpressed cofilin. Also, cell cycle analysis showed that approx 90% of cofilin overexpressing cells were arrested in G1 phase, which is consistent with previous reports that drug-mediated disruption of actin filaments can cause G I phase arrest. Taken together, cofilin overexpression cell model provides evidence that the effects of actin cytoskeletal destabilization on cell cycle progression can be studied using molecular approach instead of drug.