The Notch signaling pathway is related to neurovascular progression of pancreatic cancer
The Notch signaling pathway is related to neurovascular progression of pancreatic cancer
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DOI：
10.1097/01.sla.0000189115.94847.f1
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发表时间：
2005-12-01
影响因子：
9
通讯作者：
Friess, H
中科院分区：
文献类型：
--
作者：
Büchler, P;Gazdhar, A;Friess, H
目的：分析Notch信号通路在胰腺癌血管生成和侵袭中的潜在作用。背景：血管生成、疼痛和早期神经侵袭是胰腺癌的临床特征。血管和神经一起发育，并在生物体中使用共同的路线。 Notch 通路（Notch-1/4、Jagged-1/2、Delta-1）在此过程中显得至关重要。本研究分析了胰腺癌中的Notch通路，并表征了其血管生成和侵袭作用。方法：培养5个PaCa细胞系进行体外实验。采用实时定量 RT-PCR 来量化 31 份人类 PaCa 样本中的 mRNA 表达，并使用免疫组织化学来定位肿瘤样本中的蛋白质表达。 Notch 信号传导的激活是通过用组成型活性 Notch-1 突变体 (Notch-IC) 转染 PaCa 细胞来完成的。 Jagged 和 Delta 的过表达是通过全长 cDNA 的转染实现的。使用球体测定来研究血管生成，并使用 ELISA 来测量 VEGF、bFGF 和血管生成素的表达。采用Matrigel侵袭实验分析肿瘤细胞侵袭能力。结果：Notch-3和Notch-4 mRNA在PaCa中显着过表达（P < 0.001）。免疫组织化学也揭示了 Notch-1 的蛋白质积累。所有配体均显着上调。在神经、血管和导管肿瘤细胞中观察到配体的阳性免疫信号。用组成型活性Notch-IC突变体和Jagged-1转染PaCa细胞显示VEGF水平增加。与此同时，重组 Jagged-1 在球体测定中增加了内皮细胞的出芽。结论：Notch 通路最有可能调节胰腺癌中的神经血管发育。组成型 Notch-1 突变体和 Jagged-1 激活该信号通路会导致所有血管生成和侵袭性肿瘤表型。因此，特异性阻断 Notch 信号传导可能对胰腺癌患者有益。
Objective: To analyze the potential role of the Notch signaling pathway in pancreatic cancer angiogenesis and invasion.Background: Angiogenesis, pain, and early neuroinvasion are clinical features of pancreatic cancer. Blood vessels and nerves develop together and use common routes through the organism. The Notch pathway (Notch-1/4, Jagged-1/2, Delta-1) appears crucial in this process. The current study analyzed the Notch pathway in pancreatic cancer and characterized its angiogenic and invasive effects.Methods: Five PaCa cell lines were cultured for the in vitro experiments. Real-time quantitative RT-PCR was done to quantify mRNA expression in 31 human PaCa specimens, and immunohistochemistry was used to localize protein expression within tumor specimens. Activation of the Notch signaling was done by transfection of PaCa cells with a constitutive active Notch-1 mutant (Notch-IC). Overexpression of Jagged and Delta was achieved by transfection of full-length cDNA. Spheroid assays were used to study angiogenesis and ELISAs to measure VEGF, bFGF, and angiogenin expression. Matrigel invasion assays were used to analyze tumor cell invasion.Results: Notch-3 and Notch-4 mRNA were significantly (P < 0.001) overexpressed in PaCa. Immunohistochemistry revealed protein accumulation of Notch-1 as well. All ligands were significantly up-regulated. A positive immunosignal of ligands was seen in nerves, blood vessels, and ductal tumor cells. Transfection of PaCa cells with the constitutive active Notch-IC mutant and with Jagged-1 revealed increased levels for VEGF. Concomitantly, recombinant Jagged-1 increased sprouting of endothelial cells in the spheroid assay.Conclusion: The Notch pathway most likely regulates neurovascular development in pancreatic cancer. Activation of this signaling pathway by constitutive Notch-1 mutants and by Jagged-1 causes all angiogenic and invasive tumor phenotype. Specific blockade of Notch signaling may therefore be beneficial for patients with pancreatic cancer.