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

Background: Adenoid cystic carcinoma is a high malignant carcinoma characterized by intensive local invasion and high incidence of distant metastasis. Although many reports have demonstrated that angiogenesis has played an important role in tumor metastasis, the relationship between metastasis characters and angiogenesis ability in high and low metastasis cell lines of Adenoid cystic carcinoma has rarely been reported. The present study aimed to compare the angiogenesis ability of ACC-M ( high metastasis) and ACC-2 ( low metastasis) cell lines in vitro. Furthermore, the activity of nuclear factor kappa B and the expression of vascular endothelial growth factor ( VEGF) in ACC-2 and ACC-M were also detected. Methods: Electrophoretic mobility shift assay was used to detect nuclear factor kappa 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 kappa 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-I kappa B alpha M vector into ACCs cell lines expressing the phosphorylation defective I kappa B alpha M. Results: Nuclear factor kappa B activity and the rate of nuclear factor kappa 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 kappa 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. Conclusion: These results implicated that ACCs cells with higher metastasis feature might present greater angiogenesis ability.