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BMC Cancer  2007 

In vitro angiogenesis and expression of nuclear factor κB and VEGF in high and low metastasis cell lines of salivary gland Adenoid Cystic Carcinoma

DOI: 10.1186/1471-2407-7-95

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Electrophoretic mobility shift assay was used to detect nuclear factor κappa B activity. Semi-quantitative RT-PCR was used to quantify the mRNA level of VEGF. Immuofluorescence double staining and semi-quantitative confocal laser scanning analysis was carried out to detect nuclear factor κappa B nuclear localization and staining intensity of VEGF. The angiogenesis ability of ACC-M and ACC-2 was compared by an in vitro three-dimensional angiogenic model assay. The vector transfection assay was performed to transfect the PCMV-IκBαM vector into ACCs cell lines expressing the phosphorylation defective IκBαM.Nuclear factor κappa B activity and the rate of nuclear factor κappa B nuclear localization in ACC-M was significantly higher than that in ACC-2. Moreover, ACC-M exhibited higher mRNA and protein levels of vascular endothelial growth factor than ACC-2. VEGF mRNA expression was effectively decreased by inhibition of nuclear factor κappa B activity. Furthermore, ACC-M could remarkably stimulate the migration and tube formation of endothelial cells and induce The umbilical vein endothelial cells sprouting into the gel matrix.These results implicated that ACCs cells with higher metastasis feature might present greater angiogenesis ability.Adenoid cystic carcinoma of salivary glands (ACCs) is a high malignant carcinoma characterized by intensive local invasion and insidious distant metastasis to the lung at an early stage, which is responsible for a poor long-term survival rate [1]. Although reasons of the invasiveness and aggressive metastatic dissemination of ACCs remain unclear, angiogenesis might be a possible involving mechanism [2]. Our previous study showed that the ACCs histological type which presented high metastasis tendency exhibited higher microvessel density levels and more intensive expression of angiogenic related factors [3].Angiogenesis, the development of new blood vessels from the pre-existing vascular beds, is an essential pathophysiologic event occur


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