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通过敲低髓母细胞瘤细胞中 Rac1 的表达来抑制肿瘤细胞的迁移和侵袭。

Inhibition of tumor cell migration and invasion through knockdown of Rac1 expression in medulloblastoma cells.

机构信息

Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, Guangzhou, China.

出版信息

Cell Mol Neurobiol. 2011 Mar;31(2):251-7. doi: 10.1007/s10571-010-9615-8. Epub 2010 Nov 13.

Abstract

Medulloblastoma is one of the common malignant brain tumors in children or young adults and its overall disease-free 5-year survival rate is approximately 50% due to tumor progression, invasion, and metastasis. This study aimed to determine whether one of Rho GTPases, Rac1 can affect the morphology, motility, and invasion of medulloblastoma cells through knockdown of Rac1 expression. Medulloblastoma Daoy cells were used to manipulate Rac1 expression using Rac1 shRNA, Rac1N17, and Rac1L61 constructs. Reverse transcriptase-PCR and western blot were used to detect expression of Rac1 mRNA and protein, respectively. Invasion and migration assays were performed to assess invasion and migration capacity of Daoy cells, respectively. The data showed that Rac1 mRNA and protein were overexpressed in Daoy cells. Deletion of Rac1 decreased the cross-linked actin network and pseudopodia and also inhibited the number of migration cells migrated or invaded to the other side of the filter compared to control cells. These data indicated that the invasion and migration in Daoy cells were inhibited by deletion of Rac1, and suggest that targeting Rac1 by Rac1 shRNA may further be evaluated and used as a potential anticancer strategy to treat medulloblastoma.

摘要

髓母细胞瘤是儿童或青少年常见的恶性脑肿瘤之一,由于肿瘤的进展、侵袭和转移,其总无病 5 年生存率约为 50%。本研究旨在通过敲低 Rac1 表达,确定 Rho GTPases 之一 Rac1 是否会影响髓母细胞瘤细胞的形态、运动和侵袭。本研究使用 Rac1 shRNA、Rac1N17 和 Rac1L61 构建体来操纵髓母细胞瘤 Daoy 细胞中的 Rac1 表达。逆转录 -PCR 和 Western blot 分别用于检测 Rac1 mRNA 和蛋白的表达。侵袭和迁移实验分别用于评估 Daoy 细胞的侵袭和迁移能力。结果表明,Rac1 mRNA 和蛋白在 Daoy 细胞中过表达。与对照组细胞相比,Rac1 的缺失减少了交联的肌动蛋白网络和伪足,并且抑制了迁移细胞迁移或侵袭到滤器另一侧的数量。这些数据表明,Rac1 的缺失抑制了 Daoy 细胞的侵袭和迁移,提示通过 Rac1 shRNA 靶向 Rac1 可能进一步被评估并用作治疗髓母细胞瘤的潜在抗癌策略。

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本文引用的文献

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Plasticity of cell migration: a multiscale tuning model.细胞迁移的可塑性:一种多尺度调谐模型。
J Cell Biol. 2010 Jan 11;188(1):11-9. doi: 10.1083/jcb.200909003. Epub 2009 Dec 1.
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Coordination of Rho GTPase activities during cell protrusion.细胞突起过程中Rho GTP酶活性的协调。
Nature. 2009 Sep 3;461(7260):99-103. doi: 10.1038/nature08242. Epub 2009 Aug 19.
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Mechanical modes of 'amoeboid' cell migration.“阿米巴样”细胞迁移的机械模式。
Curr Opin Cell Biol. 2009 Oct;21(5):636-44. doi: 10.1016/j.ceb.2009.05.003. Epub 2009 Jun 11.
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Rho GTPases in cancer cell biology.Rho GTP酶在癌细胞生物学中的作用
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