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Abstract：
Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive neoplastic diseases, associated with a remarkably poor prognosis. However, the molecular mechanisms underlying the development of PDAC remain elusive. The aim of this study was to identify genes whose expressions are correlated with a poor prognosis in PDAC patients, and to unravel the mechanisms underlying the involvement of these genes in the development of the cancer.
Global gene expression profiling was conducted in 39 specimens obtained from Japanese patients with PDAC to identify genes whose expressions were correlated with a shorter overall survival. The effect of gene silencing or overexpression of ARHGEF15 in pancreatic cancer cell lines was examined by introducing siRNAs of ARHGEF15 or the ARHGEF15 expression vector. After assessing the effect of ARHGEF15 deregulation on the Rho-family proteins by pull-down assay, wound healing, transwell and cell viability assays were carried out to investigate the cellular phenotypes caused by the perturbation.
The global mRNA expression profiling revealed that overexpression of ARHGEF15, a Rho-specific GEF, was significantly associated with a poor prognosis in patients with PDAC. We also found that the depletion of ARHGEF15 by RNA interference in pancreatic cancer cell lines downregulated the activities of molecules of the Rho signaling pathway, including RhoA, Cdc42 and Rac1. Then, we also showed that ARHGEF15 silencing significantly reduced the motility and viability of the cells, while its overexpression resulted in the development of the opposite phenotype in multiple pancreatic cancer cell lines.
These data suggest that upregulation of ARHGEF15 contributes to the development of aggressive PDAC by increasing the growth and motility of the pancreatic cancer cells, thereby worsening the prognosis of these patients. Therefore, ARHGEF15 could serve as a novel therapeutic target in patients with PDAC.
Global gene expression profiling was conducted in 39 specimens obtained from Japanese patients with PDAC to identify genes whose expressions were correlated with a shorter overall survival. The effect of gene silencing or overexpression of ARHGEF15 in pancreatic cancer cell lines was examined by introducing siRNAs of ARHGEF15 or the ARHGEF15 expression vector. After assessing the effect of ARHGEF15 deregulation on the Rho-family proteins by pull-down assay, wound healing, transwell and cell viability assays were carried out to investigate the cellular phenotypes caused by the perturbation.
The global mRNA expression profiling revealed that overexpression of ARHGEF15, a Rho-specific GEF, was significantly associated with a poor prognosis in patients with PDAC. We also found that the depletion of ARHGEF15 by RNA interference in pancreatic cancer cell lines downregulated the activities of molecules of the Rho signaling pathway, including RhoA, Cdc42 and Rac1. Then, we also showed that ARHGEF15 silencing significantly reduced the motility and viability of the cells, while its overexpression resulted in the development of the opposite phenotype in multiple pancreatic cancer cell lines.
These data suggest that upregulation of ARHGEF15 contributes to the development of aggressive PDAC by increasing the growth and motility of the pancreatic cancer cells, thereby worsening the prognosis of these patients. Therefore, ARHGEF15 could serve as a novel therapeutic target in patients with PDAC.
摘要：
胰腺导管腺癌(PDAC)是侵袭性最强的肿瘤性疾病之一。与显著不良预后相关。然而,PDAC发展的分子机制仍然难以捉摸。这项研究的目的是确定表达与PDAC患者预后不良相关的基因。并阐明这些基因参与癌症发展的潜在机制。
在从日本PDAC患者获得的39个样本中进行了全局基因表达谱分析，以鉴定其表达与较短的总生存期相关的基因。通过引入ARHGEF15或ARHGEF15表达载体的siRNA来检查ARHGEF15基因沉默或过表达在胰腺癌细胞系中的作用。在通过下拉法评估ARHGEF15失调对Rho家族蛋白的影响后，伤口愈合，进行了transwell和细胞活力测定，以研究由扰动引起的细胞表型。
全局mRNA表达谱显示，Rho特异性GEFARHGEF15的过表达，与PDAC患者的不良预后显著相关。我们还发现，在胰腺癌细胞系中通过RNA干扰清除ARHGEF15下调了Rho信号通路分子的活性，包括RhoA,Cdc42和Rac1。然后，我们还表明，ARHGEF15沉默显着降低了细胞的运动性和活力，而其过表达导致多种胰腺癌细胞系中相反表型的发展。
这些数据表明，ARHGEF15的上调通过增加胰腺癌细胞的生长和运动来促进侵袭性PDAC的发展。从而使这些患者的预后恶化。因此,ARHGEF15可以作为PDAC患者的新治疗靶点。
在从日本PDAC患者获得的39个样本中进行了全局基因表达谱分析，以鉴定其表达与较短的总生存期相关的基因。通过引入ARHGEF15或ARHGEF15表达载体的siRNA来检查ARHGEF15基因沉默或过表达在胰腺癌细胞系中的作用。在通过下拉法评估ARHGEF15失调对Rho家族蛋白的影响后，伤口愈合，进行了transwell和细胞活力测定，以研究由扰动引起的细胞表型。
全局mRNA表达谱显示，Rho特异性GEFARHGEF15的过表达，与PDAC患者的不良预后显著相关。我们还发现，在胰腺癌细胞系中通过RNA干扰清除ARHGEF15下调了Rho信号通路分子的活性，包括RhoA,Cdc42和Rac1。然后，我们还表明，ARHGEF15沉默显着降低了细胞的运动性和活力，而其过表达导致多种胰腺癌细胞系中相反表型的发展。
这些数据表明，ARHGEF15的上调通过增加胰腺癌细胞的生长和运动来促进侵袭性PDAC的发展。从而使这些患者的预后恶化。因此,ARHGEF15可以作为PDAC患者的新治疗靶点。
