Identification of genes regulating migration and invasion using a new model of metastatic prostate cancer.
Identification of genes regulating migration and invasion using a new model of metastatic prostate cancer.
复制标题
DOI：
10.1186/1471-2407-14-387
复制
发表时间：
2014-05-30
期刊：
影响因子：
3.8
通讯作者：
Bielenberg DR
中科院分区：
文献类型：
--
作者：
Banyard J;Chung I;Migliozzi M;Phan DT;Wilson AM;Zetter BR;Bielenberg DR
了解复杂的、多步骤的转移过程仍然是癌症研究的主要挑战。转移模型可以揭示肿瘤的发展和进展，并为测试新的干预策略提供工具。为了建立新的肿瘤转移模型，我们使用了DU145人前列腺癌细胞，并进行了反复的原位前列腺注射和后续淋巴结转移的选择。分析肿瘤生长、转移、细胞迁移及侵袭情况。微阵列分析用于鉴定与转移相关的细胞迁移和癌症相关基因。选择的基因用siRNA沉默，并在transwell迁移和Matrigel入侵实验中确定它们在细胞迁移和侵袭中的作用。我们的体内循环策略创造了具有显著增加肿瘤发生和增加淋巴结定植能力的细胞系（DU145LN1-LN4）。前列腺肿瘤异种移植显示血管化增加，podoplanin阳性淋巴管增大，边缘浸润。微阵列分析揭示了与转移潜能相关的基因表达谱。通过基因网络分析，我们选择了3个显著上调的细胞运动和癌症相关基因进行进一步分析：EPCAM（上皮细胞粘附分子）、ITGB4（整合素β4）和PLAU（尿激酶型纤溶酶原激活物（uPA））。与亲代DU145相比，这些基因在转移性更强的DU145- ln4细胞中的蛋白表达均有所增加。SiRNA敲低EpCAM、整合素-β4或uPA均显著降低DU145-LN4细胞的细胞迁移。相比之下，只有uPA siRNA抑制细胞对Matrigel的侵袭。使用uPA抑制剂amiloride和UK122证实了uPA在细胞侵袭中的作用。我们的方法已经确定了转移性肿瘤细胞迁移和侵袭所需的基因，我们提出我们的新的体内模型系统将成为询问前列腺癌转移级联的有力工具。
Understanding the complex, multistep process of metastasis remains a major challenge in cancer research. Metastasis models can reveal insights in tumor development and progression and provide tools to test new intervention strategies. To develop a new cancer metastasis model, we used DU145 human prostate cancer cells and performed repeated rounds of orthotopic prostate injection and selection of subsequent lymph node metastases. Tumor growth, metastasis, cell migration and invasion were analyzed. Microarray analysis was used to identify cell migration- and cancer-related genes correlating with metastasis. Selected genes were silenced using siRNA, and their roles in cell migration and invasion were determined in transwell migration and Matrigel invasion assays. Our in vivo cycling strategy created cell lines with dramatically increased tumorigenesis and increased ability to colonize lymph nodes (DU145LN1-LN4). Prostate tumor xenografts displayed increased vascularization, enlarged podoplanin-positive lymphatic vessels and invasive margins. Microarray analysis revealed gene expression profiles that correlated with metastatic potential. Using gene network analysis we selected 3 significantly upregulated cell movement and cancer related genes for further analysis: EPCAM (epithelial cell adhesion molecule), ITGB4 (integrin β4) and PLAU (urokinase-type plasminogen activator (uPA)). These genes all showed increased protein expression in the more metastatic DU145-LN4 cells compared to the parental DU145. SiRNA knockdown of EpCAM, integrin-β4 or uPA all significantly reduced cell migration in DU145-LN4 cells. In contrast, only uPA siRNA inhibited cell invasion into Matrigel. This role of uPA in cell invasion was confirmed using the uPA inhibitors, amiloride and UK122. Our approach has identified genes required for the migration and invasion of metastatic tumor cells, and we propose that our new in vivo model system will be a powerful tool to interrogate the metastatic cascade in prostate cancer.
登录
查看更多内容
影响因子：
4.6
作者：
Banyard J;Chung I;Wilson AM;Vetter G;Le Béchec A;Bielenberg DR;Zetter BR
通讯作者：
Zetter BR
影响因子：
3.7
作者：
Mamoune, A;Kassis, J;Wells, A
通讯作者：
Wells, A
影响因子：
2.4
作者：
Gerson KD;Maddula VS;Seligmann BE;Shearstone JR;Khan A;Mercurio AM
通讯作者：
Mercurio AM
DOI：
10.1016/j.urolonc.2011.03.007
发表时间：
2013-05-01
影响因子：
2.7
作者：
Benko, Goran;Spajic, Borislav;Tomas, Davor
通讯作者：
Tomas, Davor
影响因子：
4.8
作者：
Drake, Justin M.;Barnes, J. Matthew;Henry, Michael D.
通讯作者：
Henry, Michael D.