Rac1 and Cdc42 are regulators of HRasV12-transformation and angiogenic factors in human fibroblasts.
Rac1 and Cdc42 are regulators of HRasV12-transformation and angiogenic factors in human fibroblasts.
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DOI：
10.1186/1471-2407-10-13
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发表时间：
2010-01-12
期刊：
影响因子：
3.8
通讯作者：
McCormick JJ
中科院分区：
文献类型：
--
作者：
Appledorn DM;Dao KH;O'Reilly S;Maher VM;McCormick JJ
Rac1 和 Cdc42 的活性对于 HRas 诱导的啮齿动物成纤维细胞转化至关重要。更重要的是，Rac1和/或Cdc42的组成型激活突变体的表达足以使其恶性转化。尚未研究这两种 Rho GTP 酶在 HRas 介导的人类成纤维细胞转化中的作用。在这里，我们评估了 Rac1 和 Cdc42 对维持 HRas 诱导的人成纤维细胞转化的贡献，并确定了 Rac1 或 Cdc42 组成型激活突变体诱导人成纤维细胞株恶性转化的能力。在四环素可调节启动子的控制下，Rac1 和 Cdc42 的显性失活突变体在人 HRas 转化的肿瘤来源的成纤维细胞系中表达。这些细胞用于确定 Rac1 和/或 Cdc42 蛋白在维持 HRas 诱导的转化表型中的作用。类似地，在未转化的人成纤维细胞株中表达组成型活性突变体，以评估它们诱导恶性转化的潜力。 Affymetrix GeneChip 阵列用于转录组分析，随后使用蛋白质测定验证观察到的表达差异。显性失活 Rac1 和/或 Cdc42 的表达显着改变了 HRas 恶性转化的人成纤维细胞的转化表型。相反，Rac1或Cdc42的组成型活性突变体的表达不足以诱导恶性转化。微阵列分析显示，29 个基因的表达依赖于 Rac1 和 Cdc42，其中许多基因已知在癌症中发挥作用。 uPA 和 VEGF 这两个基因的依赖性在常氧和缺氧条件下得到了进一步验证。这里提出的结果表明，Rac1 和 Cdc42 的表达对于维持致癌 HRas 转化的人类细胞中的几种转化表型是必要的，包括它们在无胸腺小鼠中形成肿瘤的能力。我们的数据还表明，单独表达活化的 Rac1 或 Cdc42 不足以导致人类成纤维细胞的恶性转化，尽管每种表达都是特定转化表型所必需的。此外，我们的研究阐明了几个高度重要的癌症相关基因的表达需要 Rac1 和/或 Cdc42 的活性，这也可能在细胞转化中发挥关键作用。
The activities of Rac1 and Cdc42 are essential for HRas-induced transformation of rodent fibroblasts. What is more, expression of constitutively activated mutants of Rac1 and/or Cdc42 is sufficient for their malignant transformation. The role for these two Rho GTPases in HRas-mediated transformation of human fibroblasts has not been studied. Here we evaluated the contribution of Rac1 and Cdc42 to maintaining HRas-induced transformation of human fibroblasts, and determined the ability of constitutively activated mutants of Rac1 or Cdc42 to induce malignant transformation of a human fibroblast cell strain. Under the control of a tetracycline regulatable promoter, dominant negative mutants of Rac1 and Cdc42 were expressed in a human HRas-transformed, tumor derived fibroblast cell line. These cells were used to determine the roles of Rac1 and/or Cdc42 proteins in maintaining HRas-induced transformed phenotypes. Similarly, constitutively active mutants were expressed in a non-transformed human fibroblast cell strain to evaluate their potential to induce malignant transformation. Affymetrix GeneChip arrays were used for transcriptome analyses, and observed expression differences were subsequently validated using protein assays. Expression of dominant negative Rac1 and/or Cdc42 significantly altered transformed phenotypes of HRas malignantly transformed human fibroblasts. In contrast, expression of constitutively active mutants of Rac1 or Cdc42 was not sufficient to induce malignant transformation. Microarray analysis revealed that the expression of 29 genes was dependent on Rac1 and Cdc42, many of which are known to play a role in cancer. The dependence of two such genes, uPA and VEGF was further validated in both normoxic and hypoxic conditions. The results presented here indicate that expression of both Rac1 and Cdc42 is necessary for maintaining several transformed phenotypes in oncogenic HRas transformed human cells, including their ability to form tumors in athymic mice. Our data also indicate that expression of either activated Rac1 or Cdc42 alone is not sufficient for malignant transformation of human fibroblasts, although each is required for specific transformed phenotypes. Furthermore, our study elucidates that the expression of several highly significant cancer related genes require the activities of Rac1 and/or Cdc42 which may also play a critical role in cellular transformation.
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DOI：
10.1158/1541-7786.mcr-08-0577
发表时间：
2009-06
期刊：
Molecular cancer research : MCR
影响因子：
--
作者：
Matsuo Y;Campbell PM;Brekken RA;Sung B;Ouellette MM;Fleming JB;Aggarwal BB;Der CJ;Guha S
通讯作者：
Guha S
影响因子：
8
作者：
Aguirre-Ghiso, JA;Frankel, P;Foster, DA
通讯作者：
Foster, DA
影响因子：
4.8
作者：
Milanini, J;Viñals, F;Pagès, G
通讯作者：
Pagès, G
影响因子：
11.2
作者：
Holland, SJ;Powell, MJ;Lorens, JB
通讯作者：
Lorens, JB
影响因子：
5.3
作者：
Lasa, M;Mahtani, KR;Clark, AR
通讯作者：
Clark, AR