Upregulation of molecular motor‐encoding genes during hepatocyte growth factor‐and epidermal growth factor‐induced cell motility
Upregulation of molecular motor‐encoding genes during hepatocyte growth factor‐and epidermal growth factor‐induced cell motility
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DOI：
10.1002/(sici)1097-4652(199606)167:3
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发表时间：
1996-06
影响因子：
5.6
通讯作者：
N. Török;R. Urrutia;Toshikazu Nakamura;M. McNiven
中科院分区：
文献类型：
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作者：
N. Török;R. Urrutia;Toshikazu Nakamura;M. McNiven
已知肝细胞生长因子（HGF）和表皮生长因子（EGF）刺激培养物中上皮细胞的运动。然而，介导这些重要变化的分子机制知之甚少。在这里，我们已经确定了HGF和EGF对肝细胞形态，细胞骨架组织和分子马达编码基因表达的影响。原代培养的肝细胞用10 ng/ml的HGF或EGF处理，并在处理后10、24和48 h用相位和荧光显微镜观察。我们发现，随着时间的推移，处理的细胞在处理24小时后伸展并变得细长，同时在48小时前形成长突起，微管和肌动蛋白细胞骨架发生显着变化。进行定量北方印迹分析，以测量可能导致这种形式变化的细胞骨架-（β-肌动蛋白、α-微管蛋白）和分子马达-（动力蛋白、驱动蛋白和肌球蛋白Iα和II）编码基因的表达。我们观察到在HGF处理仅2小时后，肌球蛋白II（3.3倍）、驱动蛋白（2.7倍）、肌球蛋白Iα（2.2倍）和α微管蛋白（1.9倍）的表达水平增加最多。相反，EGF在10 h时上调肌球蛋白Iα（2.4倍）、驱动蛋白（1.5倍）和动力蛋白（1.5倍）的表达。β-actin基因的表达在HGF处理的细胞中保持恒定，而EGF在处理10小时后诱导轻微上调。这些结果首次表明，分子马达编码基因的选择性上调与HGF和EGF诱导的细胞形状和运动性的改变相关。© 1996 Wiley利斯公司
Hepatocyte growth factor (HGF) and epidermal growth factor (EGF) are known to stimulate the locomotion of epithelial cells in culture. However, the molecular mechanisms which mediate these important changes are poorly understood. Here we have determined the effects of HGF and EGF on hepatocyte morphology, cytoskeletal organization, and the expression of molecular motor‐encoding genes. Primary cultures of hepatocytes were treated with 10 ng/ml of HGF or EGF and observed with phase and fluorescence microscopy at 10, 24, and 48 h after treatment. We found that, over time, treated cells spread and became elongated after 24 h of treatment while forming long processes with dramatic alterations in the microtubule and actin cytoskeletons by 48 h. Quantitative Northern blot analysis was performed to measure expression of cytoskeletal‐(β‐actin, α‐tubulin) and molecular motor‐(dynein, kinesin, and myosin Iα and II) encoding genes which may contribute to this change in form. We observed the highest increase in levels of expression for myosin II (3.3‐fold), kinesin (2.7‐fold), myosin Iα (2.2‐ fold), and α‐tubulin (1.9‐fold) after only 2 h of treatment with HGF. In contrast, EGF upregulated the expression of myosin Iα (2.4‐fold), kinesin (1.5‐fold), and dynein (1.5‐fold) at 10 h. The expression of the β‐actin gene remained constant in HGF‐treated cells, while EGF induced a slight upregulation after 10 h of treatment. These results show for the first time that a selective upregulation of molecular motor‐encoding genes correlates with alterations in cell shape and motility induced by HGF and EGF. © 1996 Wiley‐Liss, Inc.