In vitro angiogenesis and expression of nuclear factor kappaB 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 kappaappa 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 kappaappa 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 kappaappa 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-IkappaBalphaM vector into ACCs cell lines expressing the phosphorylation defective IkappaBalphaM.


RESULTS
Nuclear factor kappaappa B activity and the rate of nuclear factor kappaappa 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 kappaappa 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.


CONCLUSIONS
These results implicated that ACCs cells with higher metastasis feature might present greater angiogenesis ability.
Zhang
Jiali
J
Key Lab for Oral Biomedical Engineering of Ministry of Education, School and Hospital of Stomatology, Wuhan University, China. ringsing@yahoo.com.cn <ringsing@yahoo.com.cn>
Peng
Bin
B
D003528:Carcinoma, Adenoid Cystic / Q000098:blood supply* / Q000378:metabolism / Q000473:pathology / Q000556:secondary*; D002465:Cell Movement; D015536:Down-Regulation; D005455:Fluorescent Antibody Technique; D006801:Humans; D066298:In Vitro Techniques; D009361:Neoplasm Invasiveness / Q000473:pathology; D009389:Neovascularization, Pathologic / Q000473:pathology*; D012016:Reference Values; D012468:Salivary Gland Neoplasms / Q000098:blood supply* / Q000378:metabolism / Q000473:pathology; D012680:Sensitivity and Specificity; D051996:Transcription Factor RelA / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
D051996:Transcription Factor RelA; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A