黏着斑激酶在缺氧促进肝癌细胞细胞骨架重组中的作用研究 |
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作者单位: | Department of Gastroenterology, Tongji Hospital, Tongji Medical CoUege, Huazhong University of Science and Technology, Wuhan 430030, China
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基金项目: | 国家自然科学基金,Young Research Foundation of Health Department of Hubei Province |
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摘 要: |
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关 键 词: | HepG2细胞 粘着斑激酶 缺氧诱导 细胞骨架 Western印迹 细胞转染 siRNA 细胞迁移 |
收稿时间: | 17 October 2009 |
Effect of focal adhesion kinase on cytoskeletal arrangement of HepG2 cells induced by hypoxia |
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Authors: | Wei Yan Yu Fu Jiazhi Liao Limin Xia Min Luo Qian Zhu Dean Tian |
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Institution: | Department of Gastroenterology, Tongji Hospital, Tongji Medical College, Huazhong University of Science andTechnology, Wuhan 430030, China |
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Abstract: | Objective: To study focal adhesion kinase (FAK) expression in hypoxic HepG2 cells and the effect of FAK siRNA on cytoskeletal arrangement of HepG2 cells induced by hypoxia. Methods: HepG2 cells were cultured in 21% O2 and 1%O2. Morphological changes were observed after hypoxia treatment. Westem blot was used to measure FAK expression. The siRNA expression vector pshRNA-FAK targeting the mRNA of FAK and vector pGensil-2 (as a control) were constructed, and then transfected into HepG2 cells. Western blot was used to detect FAK. The cytoskeletal arrangement of HepG2 cells trans fected with pshRNA-FAK induced by hypoxia was analyzed by phalloidin. The migratory ability of HepG2 cells transfected with pshRNA-FAK induced by hypoxia was analyzed by cell migration assay. Results: Hypoxia-treated cells displayed a more elongated shape with a large degree of cell detachment. FAK expression increased in hypoxic HepG2 cells. FAK protein level was decreased by 75.64% ± 3.12% (P < 0.01) after the pshRNA-FAK transfection. Hypoxia induced cytoskeletal arrangement of HepG2 cells. However, cytoskeletal arrangement of HepG2 cells transfected with pshRNA-FAK induced by hypoxia was inhibited in 1% O2. As cell migration assay showed, the migrating number of HepG cells transfected with pshRNA-FAK was significantly lower than that of control (P < 0.05). Conclusion: The expression of FAK in hypoxic HCC might have a close relationship to the cytoskeletal arrangement of HepG2 cells induced by hypoxia. Up-regulation of FAK expression may be one of mechanisms of cytoskeletal arrangement and invasion of hepatocellular carcinoma induced by hypoxia. |
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Keywords: | carcinoma hepatocellular hypoxia RNA interference focal adhesion kinase (FAK) cytoskeletal arrangement |
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